Sucrose as a risk factor for cancer of the colon and rectum: a case-control study in Uruguay.
De Stefani E. Mendilaharsu M. Deneo-Pellegrini H.
Registro Nacional de Cancer, Instituto Nacional de Oncologia, Montevideo, Uruguay.
In order to examine the relationship between dietary sucrose intake and colorectal cancer, a case-control study was conducted in Uruguay in the time period 1992-1996. In all, 289 cases and 564 controls, admitted for diagnosis or treatment in the 4 major hospitals in Montevideo, were considered eligible for the study. Total sucrose intake was associated with a monotonic positive gradient of risks and the odds ratio (OR) for the uppermost quartile of intake was of 2.18 (95% confidence interval, CI, 1.35-3.51). Glucose intake was associated with a small and non-significant increase in risk (OR 1.46, 95% CI 0.76-2.82). Finally, an interaction between sucrose and protein intake was found, and the OR for high intakes of sucrose and protein was 6.07.
Correlation between 1p deletions and aneusomy in human colorectal adenomas.
Di Vinci A. Infusini E. Peveri C. Sciutto A. Geido E. Risio M. Rossini FP. Giaretti W.
Laboratory of Biophysics and Cytometry, National Institute for Cancer Research (IST), Genoa, Italy.
Human sporadic colorectal adenomas are characterized by a relatively high occurrence of aneuploidy. Similarly, 1p deletions have been reported to be an early event in colorectal tumorigenesis, while chromosome 7, 17 and 18 gain/losses were also found. The present study investigated 1p deletions, the numerical aberrations of chromosomes 1, 7, 17 and 18, and the nuclear DNA content as obtained by flow cytometry in a series of 34 human sporadic colorectal adenomas. From these adenomas, 51 intra-adenoma regions were microdissected according to 2 degrees of dysplasia and presence of foci of early cancer. Isolated epithelial nuclei were analyzed by fluorescence in situ hybridization interphase cytogenetics using centromeric probes for chromosomes 7, 17 and 18 and, in a double-target analysis, a centromeric probe for chromosome 1 simultaneously with a telomeric probe mapping to the 1p36 band. Aneuploidy incidence due to presence of numerical aberrations for at least one among the investigated chromosomes and/or abnormal flow-cytometric DNA content was 35%, while 1p deletion incidence was 38%. The correlation of 1p deletions with aneuploidy was statistically highly significant (p = 0.003), suggesting that loss of genes in this region may be implicated in chromosome instability.
Expression of Lamp-1 and Lamp-2 and their interactions with galectin-3 in human tumor cells.
Sarafian V. Jadot M. Foidart JM. Letesson JJ. Van den Brule F. Castronovo V. Wattiaux R. Coninck SW.
Department of Physiological Chemistry, Facultes Universitaires Notre Dame De la Paix, Medical Faculty, Namur, Belgium.
Lysosomal-membrane-associated glycoproteins (Lamps) 1 and 2 are rarely found on the plasma membranes of normal cells. There is evidence suggesting an increase in their cell-surface expression in tumor cells, with some data indicating that the adhesion of some cancer cells to the extracellular matrix is partly mediated by interactions between Lamps and E-selectin and between Lamps and galectins (endogenous-galactoside-binding lectins). The present study examined the expression of Lamp-1 and Lamp-2 and their interactions with galectin-3 in different human tumor cell lines. Indirect immunofluorescence staining revealed accumulation of Lamp molecules at the edges of A2058 human metastasizing melanoma cells suggesting that these glycoproteins could participate in cell adhesion. Flow cytometry showed the presence of cell-surface Lamps in A2058, HT1080 (human fibrosarcoma) and CaCo-2 (human colon-adenocarcinoma) cells. Treatment with 2 mM sodium butyrate for 24 and 48 hr resulted in a significant increase in Lamps surface expression. A strong binding of A2058 to recombinant galectin-3 was detected by FACS. The application of 2 and 5 mM butyrate for the same incubation period enhanced galectin-3 binding to Lamps-expressing cells. Our results support the idea that Lamps may be considered a new family of adhesive glycoproteins participating in the complex process of tumor invasion and metastasis.
Protoporphyrin IX accumulation in cells treated with 5-aminolevulinic acid: dependence on cell density, cell size and cell cycle.
Moan J. Bech O. Gaullier JM. Stokke T. Steen HB. Ma LW. Berg K.
Department of Biophysics, Institute for Cancer Research, Montebello, Oslo, Norway.
Human colon adenocarcinoma cells (WiDr) and Chinese hamster lung fibroblasts cells (V79) were incubated with different concentrations of 5-aminolevulinic acid (ALA), and the production of protoporphyrin IX (PpIX) was studied using several techniques. The amount of PpIX produced per cell increased with increasing ALA concentration according to different kinetics for the 2 cell lines. For both cell lines a cell density dependency of the PpIX synthesis was observed. For saturating ALA concentrations, 2-3 times more PpIX was produced per cell at a density of 5 x 10(4) than at a density of 5 x 10(3) cells/cm2. The photosensitivity of cells appeared to increase even more than the PpIX content, indicating a cooperative effect in inactivation. The PpIX production rate increased with cell size and was about 1.9 times higher for cells in the G2 + M phase than for cells in the G1 phase of the cell cycle. Neither cell size nor cell cycle distribution were significantly dependent on cell density.
Allelotype analysis of hepatocellular carcinoma.
Piao Z. Park C. Park JH. Kim H.
Department of Pathology, Yonsei University, College of Medicine, Seoul, Korea.
To elucidate the genetic events which may play important roles in hepatocarcinogenesis, we examined every non-acrocentric chromosome arm of 22 hepatocellular carcinomas (HCCs) for loss of heterozygosity (LOH) using 68 highly polymorphic microsatellite markers. Thirty-six (92%) of 39 chromosome arms showed LOH in at least one patient, however 3 chromosome arms, 2p, 2q, and 20q, did not show any LOH. High to moderate frequency of LOH (> 30% of informative cases) was observed at chromosomes 1q (68.1%), 4q (72.7%), 8p (63.6%), 8q (77.3%), 10q (33.3%), 13q (40%), 14q (46.1%), 16q (59.1%), and 17p (46.2%). Among these, LOH on chromosomes 1q and 8q have not been previously identified in HCC. Our results suggest that novel tumor suppressor genes may be involved in the development and progression of HCC.
A meta-analysis of epidemiological studies on the combined effect of hepatitis B and C virus infections in causing hepatocellular carcinoma.
Donato F. Boffetta P. Puoti M.
Cattedra di Igiene, Universita di Brescia, Italy. email@example.com
The aim of the study was to assess whether co-infection by hepatitis-B virus (HBV) and hepatitis-C virus (HCV) is associated with a higher risk of developing hepatocellular carcinoma (HCC) than each infection alone. A meta-analysis of data published up to June 1997 was performed. HBsAg and anti-HCV antibodies or HCV RNA (anti-HCV/HCV RNA) were considered as serological markers of current HBV and HCV infection respectively. A total of 32 case-control studies were suitable for a quantitative overview. The summary odds ratios (OR) were 13.7 for HBsAg positivity and 11.5 for anti-HCV/HCV RNA positivity. The OR for anti-HCV was lower among studies using second- or third-generation anti-HCV or HCV RNA (OR, 8.2) with respect to studies with first-generation anti-HCV test (OR, 19.1). When combining data from the studies with second- or third-generation anti-HCV or HCV RNA, the OR for HBsAg positivity and anti-HCV/HCV RNA negativity was 22.5 (95% confidence interval (CI), 19.5-26.0), the OR for anti-HCV/HCV RNA positivity and HBsAg negativity was 17.3 (95% CI, 13.9-21.6), and the OR for both markers positivity was 165 (95% CI: 81.2-374, based on 191 cases and 8 controls exposed). A synergism was found between HBV and HCV infections, the OR for co-infection being greater than the sum and lower than the product of those for each infection alone. The interaction was therefore negative according to the multiplicative model, providing epidemiological evidence both of an independent effect and of interference between the 2 viruses in the carcinogenic process.
Rising incidence of biliary tract cancers in Shanghai, China.
Hsing AW. Gao YT. Devesa SS. Jin F. Fraumeni JF Jr.
Division of Cancer Epidemiology and Genetics, National Cancer Institute, Bethesda, MD 20892-7368, USA. firstname.lastname@example.org
Cancers of the biliary tract, including cancers of the gallbladder, extra-hepatic bile ducts, and ampulla of Vater, are relatively uncommon malignancies. From 1972 to 1994, biliary tract cancer was the most rapidly rising malignancy in Shanghai, China, with a 119% increase in men and 124% in women. The increase in incidence was seen for all 3 subsites, both sexes, and all age groups. Future studies are needed to identify reasons for the large increases in these rates.
Establishment of primary cultures from human colonic tissue during tumor progression: vitamin-D responses and vitamin-D-receptor expression.
Tong WM. Bises G. Sheinin Y. Ellinger A. Genser D. Potzi R. Wrba F. Wenzl E. Roka R. Neuhold N. Peterlik M. Cross HS.
Institute of General and Experimental Pathology, University of Vienna Medical School, Austria.
Primary cultures derived from pre-cancerous and cancerous human colon tissue are essential for understanding normal and abnormal growth function in the large intestine. Here presented are (i) the methodology for routine establishment of primary cultures of normal, adenoma- and carcinoma-derived cells, and (ii) data for the apparently protective role of vitamin-D compounds in colon carcinogenesis. The steroid hormone 1,25-dihydroxyvitamin D3 and some non-hypercalcemic analogs reduce the high mitotic rate of adenoma cells to that of normal colonocytes. After vitamin-D treatment, tumor cells are less proliferative and differentiation is enhanced. Primary-colon-cancer cultures display a mosaic pattern of vitamin-D-receptor expression, at the mRNA level and at the protein level, with varying intensity of expression in positive cells. This suggests that, in human colorectal tumors in vivo, a large fraction of cells will respond to genomic action of vitamin-D compounds.
Promoter-specific methylation and expression alterations of igf2 and h19 are involved in human hepatoblastoma.
Li X. Kogner P. Sandstedt B. Haas OA. Ekstrom TJ.
Department of Clinical Neuroscience, Experimental Alcohol and Drug Addiction Research Section, Karolinska Hospital, Stockholm, Sweden.
In earlier studies we found up-regulation of promoter P3 transcription and total igf2 expression with concomitant down-regulation of h19 expression in human hepatoblastoma (HB). In this study, we investigated the methylation status of these 2 genes and their abnormal regulation in 7 hepatoblastomas and 5 normal counterpart tissues. A specific part of the P3 region of igf2 was de-methylated in tumors with up-regulation of the promoter activity. A site-specific DNA hypermethylation in the h19 5'-region was found in tumor tissues with concomitant down-regulation of this gene. Therefore, abnormal methylation was found to be correlated with altered regulation of igf2 and h19 expression in human hepatoblastoma and may be involved in the genesis of this tumor.
Interleukin-6 produced by pancreatic carcinoma cells enhances humoral immune responses against tumor cells: a possible event in tumor regression.
Saito K. Ishikura H. Kishimoto T. Kawarada Y. Yano T. Takahashi T. Kato H. Yoshiki T.
Department of Pathology, Hokkaido University School of Medicine, Sapporo, Japan.
Production of interleukin-6 (IL-6) by human pancreatic carcinoma cells inversely correlates with potentials for blood-borne metastasis to the liver in nude mice. IL-6 cDNA was transfected to PCI-43, one of our cultured pancreatic carcinoma cell lines that does not produce IL-6 and generates numerous metastases to the liver. An IL-6 high-producer clone (PCI-43h) generated few metastases; IL-6 production thus has a direct effect on metastasis, whereas other transfectants (PCI-43l and PCI-43n), which are IL-6 low-, and IL-6 non-producers, respectively, did generate metastases. Tumor-reactive IgG, which mediated antibody-dependent cellular cytotoxicity (ADCC) in vitro, was detected in sera from recipient nude mice inoculated with PCI-43h but not in sera from mice given PCI-43l, PCI-43n or parent PCI-43. Tumor-reactive IgM was detected in sera from all mice, irrespective of inoculated PCI-43 species, with a slight augmentation being noted in PCI-43h-inoculated nude mice. Severe combined immunodeficiency (SCID) beige mice were then used as recipients for PCI-43 species, and tumorigeneity was examined by s.c. inoculation of a suboptimal number of PCI-43 transfectants (1 x 10(6)/0.1 ml). Only PCI-43h formed palpable masses in SCID beige mice, whereas it first grew to be palpable but subsequently became not palpable in nude mice, thereby revealing a dual action of tumor-derived IL-6. Thus, tumor-derived IL-6 confers growth promotion in SCID beige mice, while the same cytokine exhibits anti-tumorigenic functions, presumably through humoral immune responses, in nude mice.
Phase-I/II radio-immunotherapy study with Iodine-131-labeled anti-CEA monoclonal antibody F6 F(ab)2 in patients with non-resectable liver metastases from colorectal cancer.
Ychou M. Pelegrin A. Faurous P. Robert B. Saccavini JC. Guerreau D. Rossi JF. Fabbro M. Buchegger F. Mach JP. Artus JC.
Val d'Aurelle-Paul Lamarque Cancer Center, Montpellier, France. MYCHOU@valdorel.fnclcc.fr
Experimental studies in nude mice with human colon-carcinoma grafts demonstrated the therapeutic efficiency of F(ab')2 fragments to carcinoembryonic antigen (CEA) labeled with a high dose of 131Iodine. A phase I/II study was designed to determine the maximum tolerated dose of 131I-labeled F(ab')2 fragments (131I-F(ab')2) from anti-CEA monoclonal antibody F6, its limiting organ toxicity and tumor uptake. Ten patients with non-resectable liver metastases from colorectal cancer (9 detected by CT scan and 1 by laparotomy) were treated with 131I-F(ab')2, doses ranging from 87 mCi to 300 mCi for the first 5 patients, with a constant 300-mCi dose for the last 5 patients. For all the patients, autologous bone marrow was harvested and stored before treatment. Circulating CEA ranged from 2 to 126 ng/ml. No severe adverse events were observed during or immediately following infusion of therapeutic doses. The 9 patients with radiologic evidence of liver metastases showed uptake of 131I-F(ab')2 in the metastases, as observed by single-photon-emission tomography. The only toxicity was hematologic, and no severe aplasia was observed when up to 250 mCi was infused. At the 300-mCi dose, 5 out of 6 patients presented grade-3 or -4 hematologic toxicity, with a nadir for neutrophils and thrombocytes ranging from 25 to 35 days after infusion. In these 5 cases, bone marrow was re-infused. No clinical complications were observed during aplasia. The tumor response could be evaluated in 9 out of 10 patients. One patient showed a partial response of one small liver metastasis (2 cm in diameter) and a stable evolution of the other metastases, 2 patients had stable disease, and 6 showed tumor progression at the time of evaluation (2 or 3 months after injection) by CT scan. This phase-I/II study demonstrated that a dose of 300 mCi of 131I-F(ab')2 from the anti-CEA Mab F6 is well tolerated with bone-marrow rescue, whereas a dose of 200 mCi can be infused without severe bone-marrow toxicity.
Deoxyadenosine metabolism in a human colon-carcinoma cell line (LoVo) in relation to its cytotoxic effect in combination with deoxycoformycin.
Bemi V. Tazzni N. Banditelli S. Giorgelli F. Pesi R. Turchi G. Mattana A. Sgarrella F. Tozzi MG. Camici M.
We have assessed the intracellular metabolism of 2'-deoxyadenosine in a human colon-carcinoma cell line (LoVo), both in the absence and in the presence of deoxycoformycin, the powerful inhibitor of adenosine deaminase. The combination of 2'-deoxyadenosine and deoxycoformycin has been reported to inhibit the growth of LoVo cells in culture. In this paper we demonstrate that the observed toxic effect is strictly dependent on cell density. In the absence of deoxycoformycin, 2'-deoxyadenosine is primarily deaminated to 2'-deoxyinosine and then converted into hypoxanthine. In the presence of the inhibitor, the deoxynucleoside, in addition to a phosphorylation process, undergoes phosphorolytic cleavage giving rise to adenine. The conversion of 2'-deoxyadenosine to adenine might represent a protective device, emerging when the activity of adenosine deaminase is reduced or inhibited. There is much evidence to indicate that the enzyme catalyzing this process may be distinct from methylthioadenosine phosphorylase and S-adenosyl homocysteine hydrolase, which are the enzymes reported to be responsible for the formation of adenine from 2'-deoxyadenosine in mammals.
Resistance of the human colon carcinoma cell line HCT-8 to methotrexate results in selection of cells with features of enterocytic differentiation.
Barbat A. Pandrea I. Cambier D. Zweibaum A. Lesuffleur T.
INSERM U178, Villejuif, France.
Results obtained previously with the human colon carcinoma cell line HT-29 have shown that the ability of the cells to develop resistance against methotrexate (MTX) or 5-fluorouracil is restricted to cells committed to differentiate. With the aim of investigating whether this observation is cell type-specific or more general, we have extended our studies to another colon cell line, HCT-8. We have compared HCT-8 parental cells and the MTX-resistant subline HCT8-MTX using transmission electron microscopy and immuno-fluorescence detection of markers of cell polarity and differentiation. Post-confluent parental HCT-8 cells appeared highly heterogeneous and occurred in clusters of piled-up cells in which the majority were unpolarized and undifferentiated, with a minority exhibiting features of enterocyte-like cells. In contrast, HCT8-MTX cells formed domes and appeared as a monolayer of polarized cells with tight junctions and a discrete apical brush border which expressed villin, dipeptidylpeptidase-IV, CEA and the epithelial mucin MUC1. Together, our results suggest that, as in HT-29 cells, induction of resistance to MTX of HCT-8 cells results in the selection of differentiated cell types.
Expression of the alpha9beta1 integrin in human colonic epithelial cells: resurgence of the fetal phenotype in a subset of colon cancers and adenocarcinoma cell lines.
Basora N. Desloges N. Chang Q. Bouatrouss Y. Gosselin J. Poisson J. Sheppard D. Beaulieu JF.
Departement d'Anatomie et de Biologie Cellulaire, Faculte de Medecine, Universite de Sherbrooke, Quebec, Canada.
Cell-matrix interactions are thought to be of critical importance in the regulation of various cell functions, including proliferation, migration and control of gene expression. The integrins, a large family of specific receptors for the macromolecules of the extracellular matrix, are important mediators of these interactions. The integrin alpha9beta1 is one of the integrins whose expression is restricted to specialized tissues. Its exact function is unknown. In the present study, we have analyzed expression of the alpha9 subunit in human colonic epithelial cells by indirect immuno-fluorescence and Western and Northern blots. In normal intact tissues, the antigen was detected at the basolateral domain of epithelial cells in colonic glands at the fetal stage but was absent in adults. Strong staining was detected constitutively in contractile cells at both stages. In adenocarcinomas, the alpha9 subunit was detected at the basolateral domain of epithelial cells in 6 of the 10 tumors tested, while a reduction of the staining was observed in the sub-epithelial myofibroblasts in parallel with peri-glandular stroma disorganization. The potential for colon adenocarcinoma cells to express the integrin alpha9 subunit was confirmed at both the protein and transcript levels in Caco-2 and T84 cell lines, 2 well-characterized cell lines known to exhibit polarization features. The 5 other cell lines tested were negative for expression of the alpha9 subunit. Taken together, our observations suggest that the alpha9 integrin subunit is subject to an onco-fetal pattern of expression in human colonic epithelium.
Characterization of glycine-N-methyltransferase-gene expression in human hepatocellular carcinoma.
Chen YM. Shiu JY. Tzeng SJ. Shih LS. Chen YJ. Lui WY. Chen PH.
Division of Preventative Medicine, Institute of Public Health, School of Medicine, National Yang-Ming University, Taipei, Taiwan, Republic of China. email@example.com
Messenger RNA differential display was used to study liver-gene expression in paired tumor and non-tumor tissues from hepatocellular carcinoma (HCC) patients. mRNA differential display and Northern-blot analyses showed that a 0.8-kb cDNA fragment was diminished or absent from the tumorous tissues of 7 HCC patients. The cDNA fragment was sequenced and found to have 98.7% nucleotide sequence homology with human glycine-N-methyltransferase cDNA (GNMT). In addition, there was no detectable level of GNMT expression in 4 human HCC cell lines, SK-Hep1, Hep 3B, HuH-7 and HA22T, examined by Northern-blot assay. A full-length GNMT cDNA clone-9-1-2 was obtained by screening a Taiwanese liver cDNA library. In comparison with the GNMT cDNA sequence reported elsewhere, clone 9-1-2 had 4 nucleotide differences resulting in 1 amino-acid change. Immunohistochemical staining with rabbit anti-recombinant GNMT serum showed that GNMT protein almost completely disappeared in liver-cancer cells, while it was abundant in the non-tumorous liver cells. Down-regulation of GNMT gene expression may be involved in the pathogenesis of liver cancer.
p53, vessel count, and vascular endothelial growth factor expression in human colon cancer.
Takahashi Y. Bucana CD. Cleary KR. Ellis LM.
Department of Surgery, Cancer Research Institute, Kanazawa University, Japan.
We previously demonstrated an association between vascular endothelial growth factor (VEGF), vessel counts and metastasis in human colon cancer specimens. Mutant p53 has been implicated in the regulation of angiogenesis. Immuno-histochemical detection of p53 protein has been associated with p53 gene mutations. We sought to determine a correlation between p53 protein detection (i.e., mutant p53), VEGF expression and vessel counts in human colon cancer. Surgical specimens from 93 patients with colon cancer were stained immuno-histochemically for p53, VEGF and factor VIII. Vessel counts were greater in metastatic tumors than in nonmetastatic tumors and adenomas, and greater in nonmetastatic tumors than in adenomas. Vessel counts were highest in tumors with the highest VEGF expression. Vessel counts and VEGF expression were greater in p53-positive tumors than in p53-negative tumors. p53 expression correlated with both VEGF expression and vessel count. The association of p53 expression with VEGF and vessel count suggests that the poor prognosis associated with p53 mutations may be due, in part, to the role of mutant p53 in promoting angiogenesis.
Over-expression of ICAM-1, VCAM-1 and ELAM-1 might influence tumor progression in colorectal cancer.
Maurer CA. Friess H. Kretschmann B. Wildi S. Muller C. Graber H. Schilling M. Buchler MW.
Department of Visceral and Transplantation Surgery, University of Bern, Inselspital, Switzerland.
Adhesion molecules might play a role in tumor progression. We investigated expression of the adhesion molecules ICAM-1, VCAM-1 and ELAM-1 in 24 primary colorectal carcinomas using immuno-histochemistry and Northern blot analysis. Normal colonic tissue from the same patients served as controls. ICAM-1 immunostaining was restricted to the intercellular matrix and vascular endothelial cells. The vast majority of normal tissue samples revealed only faint ICAM-1 immunoreactivity. However, moderate to strong immunostaining was found in 86% of cancerous sections. The ICAM-1 immunoreaction was more intense in well-differentiated carcinomas as well as in the adenomatous parts and transition zones of cancers. Similarly, the cancers exhibited markedly enhanced VCAM-1 and ELAM-1 immunostaining in the endothelial cells of small blood vessels. The intense vascular immunostaining by ICAM-1 and VCAM-1 was associated with a strong presence of CD3-positive T lymphocytes, whereas ELAM-1 immunoreactivity did not correlate with round cell infiltration. On Northern blot analysis, ICAM-1, VCAM-1 and ELAM-1 mRNA levels were increased in 67%, 57% and 63% of carcinomas, respectively, in comparison with normal tissue samples. Densitometric analysis of Northern blots revealed an increase in ICAM-1 by 2.1-fold, an increase in VCAM-1 by 3.4-fold and an increase in ELAM-1 by 2.2-fold in cancerous tissues compared to normal controls. Over-expression of ICAM-I might prevent cell-cell disruption and, hence, tumor dissemination. Furthermore, over-expression of ICAM-1 and VCAM-1, but not ELAM-1, might favor host anti-tumor defense by trafficking of lymphocytes.
MLH1 and MSH2 constitutional mutations in colorectal cancer families not meeting the standard criteria for hereditary nonpolyposis colorectal cancer.
Genuardi M. Anti M. Capozzi E. Leonardi F. Fornasarig M. Novella E. Bellacosa A. Valenti A. Gasbarrini GB. Roncucci L. Benatti P. Percesepe A. Ponz de Leon M. Coco C. de Paoli A. Valentini M. Boiocchi M. Neri G. Viel A.
Institute of Medical Genetics, Facolta di Medicina e Chirurgia A. Gemelli, Universita Cattolica del Sacro Cuore, Rome, Italy.
Genetic diagnosis of hereditary nonpolyposis colorectal cancer (HNPCC) may have a significant impact on the clinical management of patients and their at-risk relatives. At present, clinical criteria represent the simplest and most useful method for the identification of HNPCC families and for the selection of candidates for genetic testing. However, reports of mismatch repair (MMR) gene mutations in families not fulfilling the minimal diagnostic criteria point out the necessity to identify additional clinical parameters suggestive of genetic predisposition to colorectal cancer (CRC) related to MMR defects. We thus investigated a series of 32 Italian putative HNPCC individuals selected on the basis of one of the following criteria: 1) family history of CRC and/or other extracolonic tumors; 2) early-onset CRC; and 3) presence of multiple primary malignancies in the same individual. These patients were investigated for the presence of MLH1 and MSH2 mutations by single-strand conformation polymorphism analysis. Pathogenetic truncating mutations were identified in 4 (12.5%) cases, 3 of them involving MSH2 and 1 MLH1. In addition, 2 missense MLH1 variants of uncertain significance were observed. All pathogenetic mutations were associated with early age (
Comparative analysis of histology, DNA content, p53 and Ki-ras mutations in colectomy specimens with long-standing ulcerative colitis.
Holzmann K. Klump B. Borchard F. Hsieh CJ. Kuhn A. Gaco V. Gregor M. Porschen R.
Department of Gastroenterology, University of Tubingen, Germany. firstname.lastname@example.org
Neoplastic progression in patients with chronic ulcerative colitis is characterized by the development of epithelial dysplasia, which is accompanied by genetic alterations. This study determined the time of onset of p53 and Ki-ras mutations as well as DNA aneuploidy during histological progression towards carcinoma. In all, 278 samples of 7 colectomy specimens were analyzed by flow cytometry, histology and single-strand conformation polymorphism analysis. Of the samples, 22% (61/278) were dysplastic and 43% (122/278) aneuploid, while 25% (71/278) showed p53 and 4% (11/278) Ki-ras mutations. The correlation between aneuploid status and p53 mutations varied among the patients. A strong correlation was noticed between histological progression from low-grade dysplasia to carcinoma and p53 mutations as well as DNA aneuploidy. Ki-ras mutations were found in 40% (2/5) of the carcinomatous samples. The correlation between p53 mutations and the histological status of the samples suggest the involvement of this genetic event in the development of colon cancer in patients with ulcerative colitis. In contrast to Ki-ras mutations, the appearance of p53 mutations is an early event. Therefore p53 analysis might be helpful in the classification of indefinite dysplasia and in the identification of patients at risk for cancer development. Further studies are necessary to detect the additional genetic alterations preceding the development of DNA aneuploidy.
Diet and squamous-cell cancer of the oesophagus: a French multicentre case-control study.
Launoy G. Milan C. Day NE. Pienkowski MP. Gignoux M. Faivre J.
Registre des cancers digestifs du Calvados (CJF INSERM 9603), Caen, France.
An increasing number of reports suggest that diet has an impact on oesophageal cancer risk in Western countries, where alcohol and tobacco are held to be the major determinants of the risk. The aim of our study was to identify dietary factors influencing the risk of oesophageal cancer in France and to determine whether certain of these could explain some of the geographical variations. We conducted a multicentre case-control study in 3 regions expected to have different diet and drinking habits (Normandy, Burgundy and Midi Pyrenees). Two hundred eight cases and 399 controls, all males, were interviewed about their eating, drinking and smoking habits. After proper adjustment for drinking and smoking, high consumption of butter and low consumption of fresh fish, vegetables and fruits were associated strongly and independently with an increase in oesophageal-cancer risk. Consistently, cholesterol appeared as a risk factor and vitamin E, vitamin D and phosphorus as independent protective factors. The protective effect of citrus and other fresh fruits (vitamin C) was confined strictly to heavy drinkers. Our findings suggest that more than one-third of the high incidence of oesophageal cancer in northwest France could be explained by the local excess in butter consumption, whereas geographical variations in consumption of dietary protective factors could explain no more than 10% of it. Overall, a large proportion (57%) of the excess incidence of oesophageal cancer in northwest France could be explained by local dietary habits, e.g., drinking hot Calvados liquor and excessive consumption of butter.
Protective effect of fruits and vegetables on stomach cancer in a cohort of Swedish twins.
Terry P. Nyren O. Yuen J.
Department of Medical Epidemiology, Karolinska Institutet, Stockholm, Sweden. pdt1@Columbia.edu
Observational studies, primarily of a case-control design, have shown an inverse association of fruit and vegetable consumption with the risk of stomach cancer, a finding tentatively attributed to anti-oxidant vitamins. Ensuing randomized-intervention trials of these vitamins, however, have been mostly negative. Therefore, the seemingly protective effect of fruit and vegetables in case-control studies is suspected to be influenced by the information bias inherent in the retrospective assessment of exposure, particularly since pre-conceptions about the wholesome effects of these foods are common among the public. Our aim was to examine the association of fruit and vegetable intake with the risk of stomach cancer in a prospective cohort study. Fruit and vegetable consumption was assessed in 1967 in 11,546 individuals in the Swedish Twin Registry, along with a wide range of potentially confounding factors. Complete follow-up through 1992 was attained through record linkage to the National Cancer and Death Registers. The relative risk of stomach cancer was estimated in proportional hazards models, with confidence intervals (CIs) adjusted for correlated outcomes. The risk of stomach cancer was inversely related to fruit and vegetable consumption. Controlling for potentially confounding factors, the relative risk among subjects with the lowest compared to those with the highest intake was 5.5 (95% CI 1.7-18.3) with a statistically significant dose-risk trend (p < 0.05). Our results indicate that information bias is not likely to explain the discrepancy between the results of observational studies and of randomized-intervention trials.
De novo expression of the alpha5beta1-fibronectin receptor in HT29 colon-cancer cells reduces activity of C-SRC. Increase of C-SRC activity by attachment on fibronectin.
Schmidt R. Streit M. Kaiser R. Herzberg F. Schirner M. Schramm K. Kaufmann C. Henneken M. Schafer-Korting M. Thiel E. Kreuser ED.
Department of Pharmacy, Free University of Berlin, Germany.
Changes in integrin expression during malignant transformation have been observed in many tumors. Colon-carcinoma cells show reduced expression or even loss of the alpha5beta1 integrin compared to normal or adenoma cells. To determine the significance of absent alpha5beta1 integrin signaling, we transfected the cDNA coding for the alpha5 integrin sub-unit into the human colon-carcinoma cell line HT29, which constitutively lacks this subunit but does express the beta1 subunit. We show here that the newly expressed fibronectin receptor alpha5beta1 generates multiple signals, causing marked changes in cytoskeletal arrangements within a few minutes of adhesion to fibronectin. Cells expressing the alpha5beta1 integrin exhibit the formation of actin stress fibers and focal adhesions, as well as the induction of tyrosine phosphorylation of several proteins, within 10 min. We identified the focal adhesion kinase pp125FAK and the cytoskeletal protein paxillin as major phosphorylation substrates in these cells. These proteins remained hypophosphorylated when alpha5-negative control cells were plated on fibronectin. The tyrosine kinase pp60c-src, regarded as central in the regulation of cellular proliferation and constitutively over-expressed in HT29 and in colon-carcinoma cells, showed reduced intrinsic kinase activity in unstimulated HT29alpha5 cells. In contrast, fibronectin-induced signaling through alpha5beta1 increased pp60c-src activity. Moreover, immunoprecipitation of pp60c-src from extracts of HT29alpha5 cells cultivated on fibronectin for 20 min revealed complex formation of pp60c-src and tyrosine-phosphorylated pp125FAK. Our data suggest that de novo expression of the alpha5beta1 integrin in HT29 colon-cancer cells restores signaling via pp125FAK and pp60c-src. Thus, loss of this receptor during malignant transformation may contribute to tumor-cell autonomy, while reduced activity of pp60c-src in HT29alpha5-cells may participate directly in growth control.
Sulindac sulfide alters the expression of cyclin proteins in HT-29 colon adenocarcinoma cells.
Qiao L. Shiff SJ. Rigas B.
Department of Medicine, New York Methodist Hospital, Brooklyn, USA.
Sulindac sulfide (SS), the active metabolite of the colon cancer chemopreventive compound sulindac, inhibits the proliferation of HT-29 colon cancer cells mainly by inducing cell quiescence. We determined by bivariate flow-cytometric analysis both the DNA and cyclin protein content of individual cells. Thus, we assessed in detail the expression of several cyclins during the cell-cycle phases and demonstrated that SS (i) decreases the expression of cyclins B1 and E and (ii) increases the expression of cyclins D1, D2 and D3, particularly in the G1 phase of the cell cycle. SS-induced apoptotic cells expressed both E- and D-type cyclins but not cyclin B1. The changes in cyclin expression combined with reduced catalytic activity of cyclin-dependent kinases could explain in molecular terms the anti-proliferative effect of SS on HT-29 colon cancer cells. These changes may contribute to the chemopreventive effect of sulindac.
Growth dependency of a new human pancreatic cancer cell line, YAPC, on autocrine interleukin-1alpha stimulation.
Yamada T. Okajima F. Adachi M. Ohwada S. Kondo Y.
Second Department of Surgery, Institute for Molecular and Cellular Regulation, Gunma University, Maebashi/Gunma, Japan. email@example.com
We established a new human pancreatic cancer cell line from the malignant ascites of a patient with pancreatic cancer and called it YAPC. Cytogenetic and morphological analysis indicated that this cell line is monoclonal and of human origin. YAPC cells grow in nude mice, resulting in the formation of a tumor with some functional characteristics of the original tumor. The cells secreted a large amount of inflammatory cytokines including interleukin-1alpha(IL-1alpha), IL-6 and IL-8 in the culture medium. Removal of serum from the culture medium did not change the growth rate of YAPC cells, but the removal of the conditioned medium arrested their proliferation under the serum-free conditions. Exogenous IL-1alpha but neither IL-6 nor IL-8 stimulated DNA synthesis of the cells and accelerated the progress of cell cycle from G1 to the S phase. Anti-IL-1alpha antibody prevented growth of the cells in a dose-dependent fashion. In this pancreatic cancer cell line cell growth is dependent on IL-1alpha in an autocrine fashion. This line may be a useful model for studying growth regulation mechanisms of pancreatic cancer.
Expression of four CEA family antigens (CEA, NCA, BGP and CGM2) in normal and cancerous gastric epithelial cells: up-regulation of BGP and CGM2 in carcinomas.
Kinugasa T. Kuroki M. Takeo H. Matsuo Y. Ohshima K. Yamashita Y. Shirakusa T. Matsuoka Y.
Department of Surgery, School of Medicine, Fukuoka University, Japan.
Four human carcinoembryonic antigen (CEA) family members, CEA (CD66e), non-specific cross-reacting antigen (NCA, CD66c), biliary glycoprotein (BGP, CD66a) and CEA gene-family member 2 (CGM2), are expressed in normal mucosal epithelia of the colon. Expression of BGP and CGM2 has recently been demonstrated to be down-regulated in colorectal adenocarcinomas. We have now investigated the expression of the 4 CEA family antigens in gastric adenocarcinoma and carcinoma cell lines in comparison with adjacent normal gastric mucosa. The transcripts of the CEA, NCA and BGP genes evaluated by reverse transcription-polymerase chain reaction were detectable at various levels in all the gastric adenocarcinoma cell lines tested, while CGM2 mRNA was detectable in the cell lines of poorly differentiated but not of well-differentiated carcinomas. The levels of CEA mRNA in normal gastric mucosa were variable but mostly increased in adenocarcinomas. The sparse expression of NCA observed in the normal tissues was markedly up-regulated in the carcinomas. In contrast to previous findings on normal and cancerous colonic tissues, the transcripts of CGM2 were totally undetectable and those of BGP were recognized only marginally, if at all, in normal gastric mucosa, while both messages were detected at significant levels in most of the gastric adenocarcinomas. This was confirmed by in situ hybridization. Our findings indicate that expression of the CEA family antigens, particularly that of BGP and CGM2, is differently regulated in epithelial cells of the colon and the stomach.
Expression of epidermal growth factor receptor in urinary bladder cancer metastases.
Bue P. Wester K. Sjostrom A. Holmberg A. Nilsson S. Carlsson J. Westlin JE. Busch C. Malmstrom PU.
Department of Urology, Uppsala University, Sweden.
Bladder cancers frequently exhibit an increased number of epidermal growth factor receptors (EGFR) in comparison to normal urothelium. The EGFR could potentially be a target for toxic conjugates. The aim of our study was to compare the expression of EGFR in metastases with concurrent or primary tumour in the urinary bladder using immunohistochemical techniques and a monoclonal antibody. Tumour material from 20 patients was investigated. The majority (13/20) of the metastases were homogeneously stained and showed a moderate to strong membranous staining for EGFR. The expression of EGFR in primary bladder tumours and metastases was similar. There was no indication that tumour tissue exposed to chemotherapy or radiation had a decreased number of EGFR. Targeting of the EGFR thus seems potentially applicable to metastatic disease.
Histo-blood group A/B antigen deletion/reduction vs. continuous expression in human tumor cells as correlated with their malignancy.
Ichikawa D. Handa K. Hakomori S.
Pacific Northwest Research Foundation, Department of Pathobiology, University of Washington, Seattle 98122, USA.
Deletion or reduction of histo-blood group A or B antigen in tumors of A or B individuals is clearly correlated with the degree of malignancy and metastatic potential in many types of human cancer. Haptotactic motility of A+H- or B+H- colonic or gastric tumor cell lines produced by transfection of A or B gene was significantly lower than that of parental A-H+ or B-H+ cells. This is ascribable to reduced function of alpha3 or alpha6/beta1 integrin receptor as we have recently shown. However, phenotypic changes resulting from gene transfection may not reflect physiological states associated with deletion or reduction vs. continuous expression of A or B antigen in tumors. We now describe the separation and phenotype characterization of A- cells from A+ tumor cell lines derived originally from colonic tumors of patients with histo-blood group A. A+ and A- populations were detected in originally A+ tumor cell lines SW480 and HT29. A- separated from A+ populations isolated from SW480 and HT29 were characterized by greatly enhanced haptotactic motility associated with reduced or deleted A expression at alpha3, alpha6, and beta1 integrin receptors which control cell motility. Nevertheless, expression of integrin receptors at the surface of A populations is the same as that for A+ populations for both SW480 and HT29 cells. Thus, A vs. H glycosylation in integrin receptors may alter their haptotactic function. Cell proliferation as reflected by 3H-thymidine incorporation was also reduced significantly in A+ as compared to A- populations. Our findings indicate that the degree of haptotactic motility and proliferation of colonic tumor cells are physiologically associated with the deletion or reduction vs. continuous expression of the histo-blood group A antigen.
No evidence of replication error phenotype in primary gastric lymphoma of mucosa-associated lymphoid tissue.
Xu WS. Chan AC. Liang R. Srivastava G.
Department of Pathology, The University of Hong Kong, China.
Replication error (RER) phenotype, caused by deficiency of DNA mismatch repair genes and revealed by widespread microsatellite instability, has been detected in subsets of a wide variety of solid tumors, but rarely in lymphomas in general. So far, the involvement of RER phenotype in the pathogenesis of gastric lymphoma of mucosa-associated lymphoid tissue (MALT) type has not been conclusively established. We therefore examined 9 microsatellite loci on 5 chromosomes [D2S123, D3S11, D3S1261, D3S1262, D3S1265, D6S262, D18S559, a CTTT(T) repeat in intron 20 of RBI gene and a CA repeat in p53 locus] in 33 cases of primary gastric MALT lymphoma for evidence of microsatellite instability by polymerase chain reaction using primers end-labeled with [gamma-33P] ATP. Although novel-length allele was observed in 7 of 33 cases (21.2%), none of these 7 cases showed changes in more than one locus. RER phenotype was scored as positive in a case when more than 1 of the 9 examined microsatellite loci showed length alterations. Accordingly, none of the 33 cases had a RER phenotype. This result suggests that the pathogenesis of gastric MALT lymphoma does not involve RER phenotype. It is consistent with the general observations in lymphomas, but is highly in contrast to a previous report showing more than 50% of MALT lymphomas with the RER phenotype.
Characterization of human colon cancer antigens recognized by autologous antibodies.
Scanlan MJ. Chen YT. Williamson B. Gure AO. Stockert E. Gordan JD. Tureci O. Sahin U. Pfreundschuh M. Old LJ.
Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, NY 10021, USA. firstname.lastname@example.org
The screening of cDNA expression libraries derived from human tumors with autologous antibody (SEREX) has proven to be a powerful method for defining the structure of tumor antigens recognized by the humoral immune system. In the present study, 48 distinct antigens (NY-CO-1-NY-CO-48) reactive with autologous IgG were identified by SEREX analysis in 4 patients with colon cancer. Sequencing analysis showed that 17 of the cDNA clones were previously uncharacterized molecules and 31 represented known gene products. The individual cDNA clones were analyzed in the following manner: a search for mutations or other structural changes; an analysis of mRNA expression in a panel of normal tissues; and a frequency analysis of the antibody response to the expressed product in the sera of colon cancer patients and normal individuals. The initial analysis showed NY-CO-13 to be a mutated version of the p53 tumor suppressor gene. Three of the 48 antigens showed a differential pattern of mRNA expression, with NY-CO-27 (galectin-4) expressed primarily in gastrointestinal tract, and NY-CO-37 and -38 showing a pattern of tissue-specific isoforms. With regard to immunogenicity, 20 of the 48 antigens were detected by allogeneic sera; 14 of these were reactive with sera from both normal donors and cancer patients, and 6 other clones (NY-CO-8, -9, -13, -16, -20 and -38) reacted exclusively with sera from colon cancer patients (ranging from 14% to 27%). Our results on colon cancer illustrate both the complexity and the potential of the SEREX approach for analysis of the humoral immune response against human cancer.
Dietary habits and stomach cancer in Shanghai, China.
Ji BT. Chow WH. Yang G. McLaughlin JK. Zheng W. Shu XO. Jin F. Gao RN. Gao YT. Fraumeni JF Jr.
Division of Cancer Epidemiology and Genetics, National Cancer Institute, Bethesda, MD, USA. email@example.com
Stomach cancer remains the second leading cancer in incidence in Shanghai, China, despite its decline over the past 2 decades. To clarify risk factors for this common malignancy, we conducted a population-based case-control study in Shanghai, China. Included in the study were 1,124 stomach cancer patients (age 20-69) newly diagnosed in 1988-1989 and 1,451 controls randomly selected among Shanghai residents. Usual adult dietary intake was assessed using a comprehensive food frequency questionnaire. Adjusted odds ratios (ORs) and 95% confidence intervals (CIs) were estimated using logistic regression models. Risks of stomach cancer were inversely associated with high consumption of several food groups, including fresh vegetables and fruits, poultry, eggs, plant oil, and some nutrients, such as protein, fat, fiber and antioxidant vitamins. By contrast, risks increased with increasing consumption of dietary carbohydrates, with odds ratios (ORs) of 1.5 (95% confidence interval [CI] 1.1-2.1) and 1.9 (95% CI 1.3-2.9) in the highest quartile of intake among men (p for trend=0.02) and women (p=0.0007), respectively. Similar increases in risk were associated with frequent intake of noodles and bread in both men (p=0.07) and women (p=0.05) after further adjustment for fiber consumption. In addition, elevated risks were associated with frequent consumption of preserved, salty or fried foods, and hot soup/porridge, and with irregular meals, speed eating and binge eating. No major differences in risk were seen according to subsite (cardia vs. non-cardia). Our findings add to the evidence that diet plays a major role in stomach cancer risk and suggest the need for further evaluation of risks associated with carbohydrates and starchy foods as well as the mechanisms involved.
Therapeutic efficacy and dose-limiting toxicity of Auger-electron vs. beta emitters in radioimmunotherapy with internalizing antibodies: evaluation of 125I- vs. 131I-labeled CO17-1A in a human colorectal cancer model.
Behr TM. Sgouros G. Vougiokas V. Memtsoudis S. Gratz S. Schmidberger H. Blumenthal RD. Goldenberg DM. Becker W.
Department of Nuclear Medicine, Georg-August-University, Gottingen, Germany. tmbehr@WisLAN1.med.uni-goettingen.de
Recent clinical results suggest that higher anti-tumor efficacy may be achieved with internalizing monoclonal antibodies (MAbs) at lower toxicity when labeled with Auger-electron, as compared to conventional beta-emitters. The aim of our study was to compare the toxicity and anti-tumor efficacy of the 125I-labeled internalizing MAb, CO17-1A, with its 131I-labeled form in a human colon cancer model in nude mice. Biodistribution studies were performed in nude mice bearing s.c. human colon cancer xenografts. For therapy, the mice were injected either with unlabeled 125I- or 131I-labeled C017-1A at equitoxic doses. Control groups were left untreated, were given a radiolabeled isotype-matched irrelevant antibody or a tumor-specific, but noninternalizing antibody. The maximum tolerated activities (MTD) of 131I-and 125I-CO17-1A without artificial support were 300 microCi and 3 mCi, respectively. Myelotoxicity was dose-limiting; bone marrow transplantation allowed for an increase of the MTD to 400 microCi of 131I-17-1A, whereas the MTD of 125I-17-1A with bone marrow support had not been reached at 5 mCi. Whereas no significant therapeutic effects were seen with unlabeled C017-1A, tumor growth was retarded with 131I-CO17-1A. With the 125I-label, however, therapeutic results were clearly superior. In contrast, no significant difference was observed in the therapeutic efficacy of the 131I- vs. 125I-labeled, noninternalizing antibodies. Our data indicate a superiority of Auger-electron emitters, such as 125I, as compared to therapy with conventional beta-emitters with internalizing antibodies. The lower toxicity of Auger emitters may be due to the short path length of their low-energy electrons, which can reach the nuclear DNA only if the antibody is internalized (as is the case in antigen-expressing tumor tissue, but not in the stem cells of the red marrow).
Family history and subsite of gastric cancer: data from a case-referent study in Japan.
Inoue M. Tajima K. Yamamura Y. Hamajima N. Hirose K. Kodera Y. Kito T. Tominaga S.
Division of Epidemiology, Aichi Cancer Center Research Institute, Nahoya, Japan. firstname.lastname@example.org
A comparative case-referent study was conducted using data from the Hospital-Based Epidemiologic Research Program at Aichi Cancer Center (HERPACC) (Nagoya, Japan), with the aim of clarifying the effect of family history on gastric cancer by subsite. Our study comprised 995 histologically confirmed gastric cancer cases (180 cardia, 430 middle, 365 antrum and 20 unclassified) and a total of 43,846 non-cancer outpatients at Aichi Cancer Center Hospital between 1988 and 1995. Logistic regression was used to calculate odds ratios (ORs) for family history of gastric cancer and other cancers, adjusted for age, year and season at first hospital visit, habitual smoking, habitual alcohol drinking, regular physical exercise, preference for salty food and raw vegetable intake. In both genders, a positive family history of gastric cancer was associated with a moderate, but statistically significant increase in risk of gastric cancer [OR = 1.51, 95% confidence interval (95% CI) = 1.29-1.76], while no association was observed between the risk of gastric cancer and a family history of other cancers [OR = 0.97, 95% CI = 0.84-1.13]. OR increased for the middle and antrum parts of gastric cancer, but an increment for the cardiac part was observed only in those with a maternal history of gastric cancer. Our results suggest that the risk of gastric cancer in relation to family history varies by subsite and, furthermore, that the subsite-specific risk of gastric cancer is linked to a maternal history of gastric cancer.
Matrilysin-specific antisense oligonucleotide inhibits liver metastasis of human colon cancer cells in a nude mouse model.
Hasegawa S. Koshikawa N. Momiyama N. Moriyama K. Ichikawa Y. Ishikawa T. Mitsuhashi M. Shimada H. Miyazaki K.
Kihara Institute for Biological Research, Yokohama City University, Kanagawa, Japan.
Human colon cancer frequently develops liver metastasis. Matrilysin (MMP-7), the smallest member of the matrix metalloproteinase (MMP) family, is commonly produced by human colon carcinoma cells and has been suggested to be involved in the progression and metastasis of this type of cancer. In the present study, we tested the effect of a matrilysin-specific antisense phosphorothioate oligonucleotide on liver metastasis of the human colon carcinoma cell line WiDr in nude mice. In culture, the antisense oligonucleotide moderately inhibited the secretion of matrilysin by WiDr cells. Injection of WiDr cells into the spleen of nude mice produced many metastatic tumor nodules in the liver. When the antisense oligonucleotide was injected daily into the mice for 11 days, the formation of the metastatic tumor nodules was strongly inhibited in a dose-dependent manner. An inhibition of liver metastasis of over 70% was obtained at a dose of 120 micrograms of the oligonucleotide per mouse. The antisense oligonucleotide did not inhibit tumor growth in spleen and in liver. A scrambled control oligonucleotide had no effect on liver metastasis of WiDr cells. Our results demonstrate an important role of matrilysin in liver metastasis of human colon cancer and the therapeutic potential of matrilysin antisense oligonucleotides for the prevention of metastasis.
Long circulating half-life and high tumor selectivity of the photosensitizer meta-tetrahydroxyphenylchlorin conjugated to polyethylene glycol in nude mice grafted with a human colon carcinoma.
Westerman P. Glanzmann T. Andrejevic S. Braichotte DR. Forrer M. Wagnieres GA. Monnier P. van den Bergh H. Mach JP. Folli S.
Institute of Biochemistry, University of Lausanne, Epalinges, Switzerland.
In a mode of nude mice bearing a human colon carcinoma xenograft, the biodistribution and tumor localization of metatetrahydroxyphenylchlorin (m-THPC) coupled to polyethylene glycol (PEG) were compared with those of the free form of this photosensitizer used in photodynamic therapy (PDT). At different times after i.v. injection of both forms of 125I-labeled photosensitizer, m-THPC-PEG gave on average a 2-fold higher tumor uptake than free m-THPC. In addition, at early times after injection, m-THPC-PEG showed a 2-fold longer blood circulating half-life and a 4-fold lower liver uptake than free m-THPC. The tumor to normal tissue ratios of radioactivity concentrations were always higher for m-THPC-PEG than for free m-THPC at any time point studied from 2 to 96 hr post-injection. Significant coefficients of correlation between direct fluorescence measurements and radioactivity counting were obtained within each organ tested. Fluorescence microscopy studies showed that m-THPC-PEG was preferentially localized near the tumor vessels, whereas m-THPC was more diffusely distributed inside the tumor tissue. To verify whether m-THPC-PEG conjugate remained phototoxic in vivo, PDT experiments were performed 72 hr after injection and showed that m-THPC-PEG was as potent as free m-THPC in the induction of tumor regression provided that the irradiation does for m-THPC-PEG conjugate was adapted to a well-tolerated 2-fold higher level. The overall results demonstrate first the possibility of improving the in vivo tumor localization of a hydrophobic dye used for PDT by coupling it to PEG and second that a photosensitizer conjugated to a macromolecule can remain phototoxic in vivo.
Suppression of growth of hepatocellular carcinoma by sodium butyrate in vitro and in vivo.
Yamamoto H. Fujimoto J. Okamoto E. Furuyama J. Tamaoki T. Hashimoto-Tamaoki T.
First Department of Surgery, Hyogo College of Medicine, Nishinomiya, Japan.
Treatment of HuH-7 human hepatocellular carcinoma (HCC) cells with 1-10 mM sodium butyrate (SB) resulted in growth inhibition in a dose-dependent manner. At 3 mM and higher concentrations, SB caused nuclear fragmentation and DNA ladder formation characteristic of apoptosis. In the treated cells, the expression of p21 (WAFI/CIPI) increased and that of alpha-fetoprotein (AFP) decreased. These characteristic changes were also observed with 5 other human HCC cell lines with or without mutation of the p53 gene. The ability of these cells to form colonies in soft agar was suppressed by either pretreating the cells with SB prior to soft agar plating or incubating untreated cells in SB-containing soft agar. Direct injection of SB into tumors developed from HuH-7 cells in nude mice resulted in an increase in the p21 level, a decrease in the tumor size and an increase in the survival time of mice. When the inoculation of HuH-7 cells into nude mice was immediately followed by subcutaneous injection of SB, development of tumors was either significantly delayed or completely suppressed. These results suggest that SB induces cellular differentiation and suppresses growth and tumorigenicity of HCC cells in vitro and in viva by a mechanism independent of p53 but possibly dependent on p21.
Macronutrient intake and risk of colorectal cancer in Italy.
Franceschi S. La Vecchia C. Russo A. Favero A. Negri E. Conti E. Montella M. Filiberti R. Amadori D. Decarli A.
Servizio di Epidemiologia, Centro di Riferimento Oncologico, Aviano, Italy. email@example.com
To provide further insight on the relationship between macronutrients and colorectal cancer, overall and by specific subsite(s), we carried out between 1992 and 1996 in 6 Italian areas a case-control study on 1,953 individuals of both sexes with incident colorectal cancer (age range 19-74) and 4,154 controls (age range 19-74) in hospital with acute, non-neoplastic diseases. A validated food-frequency questionnaire was used, including questions on 78 foods or recipes and on individual fat-intake pattern. The risk of cancer of the colon and rectum increased with total energy intake (odds ratio in highest vs. lowest quintile 1.43 and 1.50, respectively). The risk also rose significantly with an increase of starch intake, whereas it moderately decreased with an increase of protein intake. Monounsaturated fat intake appeared uninfluential, while saturated fat intake showed a modest direct association with rectal cancer. Polyunsaturated fat intake was inversely associated with colon cancer risk, particularly with the right colon.
Increased invasiveness of MUC1 and cDNA-transfected human gastric cancer MKN74 cells.
Suwa T. Hinoda Y. Makiguchi Y. Takahashi T. Itoh F. Adachi M. Hareyama M. Imai K.
First Department of Internal Medicine, Sapporo Medical University, Japan.
MUC1 mucin is an anti-adhesion molecule expressed in a wide variety of tumors. To examine whether MUC1 mucin is involved in tumor invasion, we have prepared MUC1 transfectants using the human gastric cancer cell line MKN74 and performed an in vivo tumor assay by transplanting these into nude mice. Tumor weight at 71 days after s.c. injection of transfectants was measured, showing that the in vivo growth of MUC1 transfectants was increased compared to that of mock transfectants. Furthermore, MUC1-transfectant tumors invaded into the muscle layer, whereas mock-transfectant tumors did not. In vitro invasion, adhesion to extracellular matrix components and phagokinetic track motility were then evaluated to analyze the mechanisms for the in vivo invasiveness of the transfectants. MUC1 transfectants exhibited an increased in vitro invasiveness, decreased binding to laminin, fibronectin, type I collogen and type IV collagen and increased motility. These effects of MUC1 mucin over-expression in MKN74 cells were abolished by the treatment of transfectants with an inhibitor of O-glycan biosynthesis, benzyl-alpha-GalNAc. Our data suggest that MUC1 mucin could be related to the increased invasive ability of MKN74 cells, whereas O-glycan might play an essential role.
Genetic instability and mutation of the TGF-beta-receptor-II gene in ampullary carcinomas.
Imai Y. Tsurutani N. Oda H. Inoue T. Ishikawa T.
Department of Pathology, Faculty of Medicine, University of Tokyo, Japan.
Ampullary carcinomas are relatively rare cancers of which very little is known in terms of carcinogenetic mechanisms at the molecular level. Genetic instability caused by mutations of mismatch-repair genes has been demonstrated to be responsible for hereditary non-polyposis colorectal cancers and a sub-set of sporadic colorectal cancers. In some of those tumors showing genetic instability, the transforming-growth-factor-beta(TGF-beta)-receptor-II gene has been found to be mutated in repetitive sequences and considered to be a target of replication error. We studied the role of genetic instability and associated TGF-beta-receptor-II-gene mutations in a series of 18 sporadic cases by analyzing 5 microsatellite loci (D2S123, D3S1029, D5S409, TP53 and BAT26) and by sequencing a poly-A repeat (nucleotides 709-718) in the TGF-beta-receptor-II gene. Microsatellite instability was observed in 4 (22.2%) and gene mutations in 14 (77.8%) cases. These data indicate that the TGF-beta-receptor-II gene might be a preferential target of genetic instability whose alteration might be specifically advantageous and constitute a common step in the development of ampullary carcinomas.
Interferon-alpha and -gamma inhibit the growth and neoplastic potential of v-src-transformed human epithelial cells by reducing Src tyrosine kinase activity.
Fujishima H. Nakano S. Tatsumoto T. Masumoto N. Niho Y.
First Department of Internal Medicine, Faculty of Medicine, Kyushu University, Fukuoka City, Japan.
To investigate whether interferons (IFNs) selectively suppress the growth of solid tumor cells with elevated protein tyrosine kinase (PTK) activity, we evaluated the effect of recombinant IFN-alpha2a and IFN-gamma on the proliferative and neoplastic potentials triggered by p60v-src using v-src-transformed HAG-1 human epithelial cells. When compared with control cells harboring the pSV2neo gene, the monolayer growth of v-src-transformed cell lines was inhibited by both recombinant IFNs, in a dose-dependent manner, whereas growth of ras-transfected cell lines was not affected. Moreover, IFNs markedly reduced the clonogenic growth of v-src-transformed cells in soft-agar rather than monolayer growth, suggesting the preferential activity of IFNs on anchorage-independent growth. Pretreatment of cells with Src or the Src-like PTK inhibitor herbimycin A or radicicol, alleviated dose-dependently the growth-inhibitory activity of IFN-alpha2a against v-src-transformed cells, suggesting that IFNs may share a common inhibitory pathway with Src PTK inhibitors. Accordingly, like herbimycin A, IFNs were found to reduce tyrosine phosphorylation of p60v-src and suppressed in vitro p60v-src kinase activity in v-src-transformed cells. Our data, together with the fact that IFNs inhibit the growth potential driven by Src but not by activated Ras, suggest that inhibition of signal transduction pathway through Src to downstream transduction events may be a primary mechanism of IFN-induced anti-prolifeative and anti-tumoral activity.
Co-expression of osteopontin and CD44v9 in gastric cancer.
Ue T. Yokozaki H. Kitadai Y. Yamamoto S. Yasui W. Ishikawa T. Tahara E.
First Department of Pathology, Hiroshima University School of Medicine, Japan.
We have examined the expression of osteopontin (OPN) in 40 human primary gastric carcinoma tissues, 5 metastatic foci (lymph nodes) and corresponding normal mucosas. Twenty-nine of 40 primary tumors (72.5%) and 3 of 5 lymph node metastases (60%) overexpressed OPN mRNA in comparison with those of the corresponding normal mucosa. The incidence as well as relative expression level of OPN mRNA was higher in well differentiated gastric cancers than poorly differentiated ones. Moreover, increased OPN mRNA expression in primary tumor specimens was observed along with the advancement of the clinico-pathological stage. Using in situ hybridization (ISH) analysis, not only inflammatory cells in tumor stroma but also tumor cells showed positive signals for OPN mRNA. By immunohistochemistry, co-immunoreaction of OPN and CD44v9 in tumor cells obviously correlated with the degree of lymphatic vessel invasion or long distant lymph node metastases in poorly differentiated gastric cancer. Interestingly, strong co-immunoreaction of OPN and CD44v9 of tumor cells was concommitant with cluster formation in the lymphatic vessels. Our results suggest that overexpression of OPN correlated with the progression of human gastric carcinoma. Especially in CD44-bearing poorly differentiated gastric cancer, interaction between OPN and CD44 may parallel lymphogenous metastasis.
Correlation of the immunohistochemical reactivity of mucin peptide cores MUC1 and MUC2 with the histopathological subtype and prognosis of gastric carcinomas.
Baldus SE. Zirbes TK. Engel S. Hanisch FG. Monig SP. Lorenzen J. Glossmann J. Fromm S. Thiele J. Pichlmaier H. Dienes HP.
Institute of Pathology, University of Cologne, Germany.
The expression of MUC1 and MUC2 mucin peptide core antigens in gastric carcinomas was studied by immunohistochemistry to determine correlations with TNM stage and histo-pathological classifications as well as a possible prognostic impact. Paraffin-embedded specimens from 128 gastric carcinomas with a minimal follow-up of 5 years were immunostained. In addition to a polyclonal antiserum generated against polymorphic epithelial mucin (MUC1) from human milk, 2 monoclonal antibodies (MAbs), HMFG2 (anti-MUC1) and 4FI (anti-MUC2), were applied. Reactivity of carcinomas was correlated with the classifications of the UICC (TNM), WHO and Lauren. Correlations with overall survival were analyzed using the Kaplan and Meier product limit method. MUC1 immunoreactivity was associated with an advanced pTNM stage. The demonstration of both mucin species (MUC1, MUC2) displayed a statistically significant correlation with tubular/papillary vs. signet-ring cell differentiation as well as with intestinal-type vs. diffuse-type of tumor growth according to Lauren. In particular, MUC2 was only rarely detectable in signet-ring cell and diffuse-type tumors. MUC1 correlated with poor prognosis in all cases and the subgroup of stage I tumors. According to the histopathological classifications, a similar result was observed in signet-ring cell and diffuse-type carcinomas. In contrast, MUC2 reactivity was associated with a favourable prognosis of intestinal-type carcinomas. In the non-neoplastic gastric mucosa, both peptide cores were recognized in the superficial epithelium, whereas parietal cells contained only MUC1, and intestinal metaplasia almost exclusively MUC2 antigens. We conclude that the mucin peptide core antigens are suitable markers for the tubule-rich gastric carcinomas, which may in part be derived from intestinal metaplasia. In addition, MUC1 may exert a prognostic relevance and appears to be involved in the progression of diffuse-type tumors.
Identification of cystatin B in human esophageal carcinoma, using differential displays in which the gene expression is related to lymph-node metastasis.
Shiraishi T. Mori M. Tanaka S. Sugimachi K. Akiyoshi T.
Department of Surgery, Medical Institute of Bioregulation, Kyushu University, Beppu, Japan.
Identification of the genes that are specifically expressed in either tumor or normal tissue is important for understanding cancer biology. Using differential displays, we obtained one gene which was specifically expressed in normal tissue but is only rarely expressed in carcinoma tissue of the human esophagus. The sequence of this gene was identical with cystatin B, known to be one of the cysteine-proteinase inhibitors, mainly inhibiting cathepsin L. There is little information on the clinical significance of cystatin B expression in human esophageal carcinoma. We thus studied the mRNA expression of cystatin B in 45 tumor/normal pair specimens of the esophagus, using the semi-quantitative reverse transcriptase polymerase chain reaction technique. The expression of cystatin B in tumor tissue was found to be markedly decreased compared with that of the corresponding normal tissue. The cases with a tumor/normal ratio of less than 0.5 showed high frequency of lymph-node metastasis and more advanced clinical stage as compared with those whose tumor/normal ratio was equal to or more than 0.5. The decreased expression of cystatin B protein in carcinoma tissue was confirmed by immunohistochemical staining. Our study suggests that reduced expression of cystatin B in esophageal-carcinoma tissue is associated with lymph-node metastasis and may therefore prove to be a useful marker for predicting the biologic aggressiveness of human esophageal carcinoma.
Involvement of vascular endothelial growth factor and urokinase-type plasminogen activator receptor in microvessel invasion in human colorectal cancers.
Nakata S. Ito K. Fujimori M. Shingu K. Kajikawa S. Adachi W. Matsuyama I. Tsuchiya S. Kuwano M. Amano J.
Department of Surgery, Shinshu University School of Medicine, Matsumoto, Nagano, Japan. firstname.lastname@example.org
To evaluate the association among known angiogenic growth factors or factors related to the plasminogen activation system and clinicopathological factors in patients with colorectal cancer, we examined the expression of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), transforming growth factor-alpha (TGF-alpha), urokinase-type plasminogen activator (u-PA), u-PA receptor (u-PA-R) and plasminogen activator inhibitor-1 (PAI-1) in clinical specimens of colorectal cancers by Northern blot analysis. In comparison with the expression of these angiogenesis-related genes in 7 paired samples of colorectal cancers and the adjacent normal mucosa, VEGF mRNA level was significantly higher in the cancer tissues than in the adjacent normal mucosa (p < 0.05). We analyzed expression of these genes in 44 cases of primary colorectal cancers. Among the 3 angiogenic growth factors we examined, VEGF mRNA expression was significantly higher in the cancer tissues with blood vessel invasion or with lymphatic vessel invasion than in those without, respectively (p < 0.05). On the other hand, u-PA-R mRNA expression was significantly higher in the cancers with blood vessel invasion than in those without (p < 0.05). In addition, there was a correlation between the expression levels of VEGF and u-PA-R mRNA in the cancer tissues we have examined. Using immunohistochemistry, strong staining of VEGF or u-PA-R was observed in the cancer cells invading the microvessels. Our findings suggest that malignant transformation might accompany the upregulation of VEGF expression in colorectal cancers and that VEGF and u-PA-R might contribute cooperatively to increase angiogenesis around the tumor as well as the metastasis via microvessels.
Expression and tissue localization of matrix metalloproteinase 7 (matrilysin) in human gastric carcinomas. Implications for vessel invasion and metastasis.
Yamashita K. Azumano I. Mai M. Okada Y.
Department of Molecular Immunology and Pathology, Cancer Research Institute, Kanazawa University, Japan.
We examined the production and tissue localization of matrix metalloproteinase-7 (MMP-7 = matrilysin) in human gastric carcinomas and analyzed the data in connection with the clinicopathological factors. Sandwich-enzyme immunoassay for the zymogen of MMP-7 (proMMP-7) showed enhanced production of MMP-7 in carcinoma tissues compared with control normal gastric mucosa. Immunohistochemical studies demonstrated that MMP-7 is localized predominantly to the carcinoma cells in 71% of the carcinoma samples (30/42 cases). The percentage of immunoreactive carcinoma cells to total carcinoma cells (positive ratio) was significantly higher in intestinal-type carcinomas (26%, median) than in diffuse-type carcinomas (3%, median) (p < 0.05). The positive ratio was markedly higher in carcinoma groups with vascular invasion (28%) or lymphatic permeation (12%) than in those without invasion (6%) or permeation (0%) (p < 0.05). It was also significantly higher in carcinoma groups with liver (49%) or lymph-node metastases (15%) than in those without metastases (6 and 2% respectively) (p < 0.05). Both proMMP-7 of 28 kDa and active MMP-7 of 19 kDa were detected in the carcinoma tissues by immunoblotting. Reverse-transcription-PCR showed specific amplification in 50% of the carcinoma cases (6/12 cases) and 8% of the normal control specimens (1/12 cases). In situ hybridization demonstrated that the carcinoma cells almost selectively express MMP-7 mRNA. These data suggest that enhanced production of proMMP-7 and its activation are implicated in invasion and metastasis of human gastric carcinomas.
Family aggregation of carcinoma of the hypopharynx and cervical esophagus: special reference to multiplicity of cancer in upper aerodigestive tract.
Morita M. Kuwano H. Nakashima T. Taketomi A. Baba H. Saito T. Tomoda H. Egashira A. Kawaguchi H. Kitamura K. Sugimachi K.
Department of Surgery II, Faculty of Medicine, Kyushu University, Fukuoka, Japan.
The role of family history in the multiple occurrence of cancer in the upper aerodigestive tract (UADT) remains unclear. The family histories of close relatives were examined in 167 patients with either hypopharyngeal or cervical esophageal cancer (PhCe cancer) and in 167 control subjects with benign diseases. The odds ratio for PhCe cancer was 2.6 in relation to family history of UADT cancers. Based on the family histories of close relatives, 167 cases with PhCe cancer were divided into 3 groups (Group I, 18 cases with a family history of UADT cancer; Group II, 37 cases with a family history of other cancers; Group III, 112 cases with no family history of any cancers). The mean age of the cases in group I was 59.4, which was younger than in group III (64.2). Second primary squamous-cell carcinomas in the UADT were more frequently recognized in group I (39%) than in group III (11%). However, no differences were observed in the smoking and drinking habits of male patients between each group. These results thus suggest that a family history of UADT cancers appears to be associated with the multiple occurrence of UADT cancers as well as the development of PhCe cancer.
Cancer incidence in Ho Chi Minh City, Viet Nam, 1995-1996.
Nguyen MQ. Nguyen CH. Parkin DM.
Ho Chi Minh Cancer Center, Binh Thanh District, Ho Chi Minh City, Viet Nam.
The results from the population-based cancer registry for the city of Ho Chi Minh in 1995-1996 represent the first information on the incidence of cancer in southern Viet Nam. A total of 4,080 cancer cases in males and 4,338 in females were registered, corresponding to age-standardized incidence rates (ASRs) of 130.9 per 100,000 in men and 100.7 per 100,000 in women. As elsewhere in South East Asia, the principal cancer of men was liver cancer (ASR 25.3), with moderately high rates of lung cancer (ASR 24.6) and stomach cancer (ASR 16.5); cancer of the penis, reportedly very common in early case series from Viet Nam, is now rarely seen. In women, cervical cancer was the dominant malignancy (ASR 26.0) followed by breast cancer (ASR 12.2) and stomach cancer (ASR 7.5). Although there may be some under-registration in these early years of operation, the recorded rates of cervical cancer and liver cancer are already high and suggest that southern Viet Nam would benefit from an effective cervical cancer screening programme, as well as efforts to interrupt the transmission of hepatitis B virus to reduce liver cancer incidence and effective anti-smoking programs.
Suppression of intracellular resistance factors by adriamycin augments heat-induced apoptosis via interleukin-1beta-converting enzyme activation in pancreatic carcinoma cells.
Kobayashi D. Watanabe N. Sasaki H. Okamoto T. Tsuji N. Sato T. Yamauchi N. Niitsu Y.
Department of Laboratory Diagnosis, Sapporo Medical University, School of Medicine, Japan.
Combination of heat and various anticancer drugs can exert a synergistic antitumor effect in vitro and in vivo, though the mechanism is not clear. We have previously shown that endogenous tumor necrosis factor (enTNF) acts as an intracellular resistance factor to inhibit the cytotoxic effect of heat by scavenging oxygen-free radicals via the induction of manganous superoxide dismutase (MnSOD). Consequently, we examined whether the suppression of these resistance factors by combining anticancer drugs and heat causes an augmentation of heat-induced cytotoxicity. The human pancreatic carcinoma cell line, PANC-1, constitutively expresses appreciable amounts of enTNF and also demonstrates heat resistance. After treatment of these cells for 15 hr with adriamycin (ADM), the expression of enTNF was decreased by 43%, and MnSOD activity was suppressed by 55%. The cytotoxic effects of the treatment of PANC-1 cells with ADM followed by heat were greater than the sum of those observed with the agents administrated individually. Heat-induced apoptosis was also augmented by pretreatment with ADM. Furthermore, the interleukin-1beta-converting enzyme inhibitor, Ac-YVAD-CMK, reversed the augmented cytotoxicity. Our results indicate that suppression of intracellular resistance factors such as enTNF and MnSOD plays an important role in apoptosis seen after heat and ADM combined therapy.
Liver metastasis and adhesion to the sinusoidal endothelium by human colon cancer cells is related to mucin carbohydrate chain length.
Bresalier RS. Byrd JC. Brodt P. Ogata S. Itzkowitz SH. Yunker CK.
Department of Medicine, Henry Ford Health Sciences Center, Detroit, MI 48202, USA. email@example.com
Mucin production by human colon cancer cells correlates with liver metastasis in animal models, but it is not known which steps in metastasis depend on specific alterations in mucin synthesis. Clonal variants of cell line LS174T selected for differences in mucin core carbohydrate expression have been further characterized biochemically, and tested for their ability to participate in metastasis-related events. LS-C mucin contains truncated carbohydrates enriched for sialyl Tn and these cells bind to basement membrane matrix to a greater extent than LS-B cells. This binding is partially inhibitable by antibody to sialyl Tn. LS-B produces more fully glycosylated mucin and preferentially binds to hepatic sinusoidal endothelial cells and E-selectin through sialylated peripheral mucin-associated carbohydrate structures. Adhesion of LS-B to endothelial cells is inhibited by neutralizing antibody to E-selectin, and inhibition of glycosylation or desialylation of LS-B mucin abrogates binding to E-selectin in vitro. LS-B cells spontaneously metastasized from cecum to liver and colonized the liver of athymic mice after splenic-portal injection to a significantly greater extent than LS-C, suggesting that expression of peripheral mucin carbohydrate structures is most important for metastasis of human colon cancer cells.