Anticancer Res

Liver metastasis of a human colorectal cancer containing two actively growing subclones.

Year 1998
Chen TR. Chang AE.
American Type Culture Collection, Rockville, MD 20852, USA.
A common pattern of karyotype evolution between clones involves gradual changes of one or only a few chromosomes. Karyotypes of one day-old cultures derived from a tumor nodule of colorectal cancer liver metastasis were studied by GTG chromosome banding. Two karyotypically distinct aneuploid CC-9-a and CC-9-b clones were found respectively, at about 1:3 ratio. Both clones showed common karyotype characteristics with the same number of copies for 13 normal and four marker chromosomes, indicating their common genetic origin. However, CC-9-b differed from CC-9-a by the loss of one copy each of eight normal and four common marker chromosomes. This mode of gross single-chromosome losses is also seen in established cell lines, and probably plays an important role in the drastic chromosome changes associated with the duplication-reduction cycle of karyotype evolution.

Tumor growth suppression by a mouse/human chimeric anti-CEA antibody and lymphokine-activated killer cells in vitro and in SCID mouse xenograft model.

Year 1998
Senba T. Kuroki M. Arakawa F. Yamamoto T. Kuwahara M. Haruno M. Ikeda S. Matsuoka Y.
First Department of Biochemistry, School of Medicine Fukuoka University, Japan.
The mouse/human chimeric antibody Ch F11-39, recently generated by ourselves, shows the same high specificity and affinity for carcinoembryonic antigen (CEA) as those of its parental mouse monoclonal antibody. Ch F11-39 is capable of mediating antibody-dependent cell-mediated cytotoxicity (ADCC) with human peripheral blood lymphocytes (PBL). Interleukin-2 (IL-2) modulates the function of immunocytes, in particular inducing lymphokine-activated killer (LAK) cells and enhancing ADCC. In the present study, we therefore tested the combination immunotherapy of Ch F11-39 with LAK cells in vitro and in severe combined immunodeficiency (SCID) mice bearing human CEA-producing tumors. In vitro experiments using human gastric tumor cell lines, Ch F11-39 effectively mediated ADCC against CEA-positive MKN-45 cells, but not against CEA-negative cells. The specificity of ADCC for Ch F11-39 was demonstrated by experiments with irrelevant target cells or irrelevant antibody. ADCC activity of PBL with Ch F11-39 was enhanced by double after preincubation with IL-2 at 10 U/ml. The concentration of Ch F11-39 required for 50% maximal cell killing was about 0.25 microgram/ml at 10 U/ml of IL-2. Increasing ADCC was triggered by IL-2 earlier (1 day) than the generation of LAK cells (3 days). Control human IgG blocked the ADCC, suggesting that the enhancement of ADCC by IL-2 may be caused by activation of effector cells expressing Fc receptors. In vivo anti-tumor activity of combined immunotherapy was estimated using SCID mice inoculated s.c. with 1 x 10(7) MKN-45 cells. The i.v. administration of LAK cells and i.p. administration of Ch F11-39 and IL-2 produced a marked growth inhibition of MKN-45 tumors in SCID mice (about 50% reduction in tumor size as compared to the control untreated group, measured 15 days after treatment). In summary, the enhanced antitumor activity of Ch F11-39 with LAK cells suggests that it might be a useful immunotherapeutic reagent for CEA-expressing tumors.

Effect of FR143430, a novel cytokine suppressive agent, on adenocarcinoma colon26-induced cachexia in mice.

Year 1998
Yamamoto N. Kawamura I. Nishigaki F. Tsujimoto S. Naoe Y. Inami M. Elizabeth L. Manda T. Shimomura K.
Exploratory Research Laboratories, Fujisawa Pharmaceutical Co., Ltd., Ibaraki, Japan.
Cancer cachexia, characterized by weight loss and progressive tissue wasting, has been postulated to be mediated by cytokines. In this study the effect of FR143430, (2-(4-fluorophenyl)-4, 5, 6, 7-tetrahydro-3-(4-pyridyl)pyrazolo[1, 5-a]pyrimidine monohydrochloride), an inhibitor of Interleukin-1 and Tumor necrosis factor-a (TNF- a), on adenocarcinoma colon26-induced cachexia was investigated in mice. Tumor growth was not affected. Nevertheless, treatment with FR143430 (0.1 to lmg) into the tumor resulted in the attenuation of the reduction in body weight, food intake, epididymal fat and carcass weight, the decrease in the circulating levels of triglyceride and glucose, and the increase in the circulating levels of total cholesterol, non esterified free fatty acid (NEFA) and total protein, which were induced by the presence of the tumor. However, oral treatment with FR143430 failed to show an inhibitory effect on cachexia induction. Overall, this study demonstrated that the cachexia induced by colon26 was alleviated by the injection of FR143430 into the tumor in sufficient quantity, without any effect on tumor growth, suggesting the potential utility of cytokine suppressive agents e for the treatment of cancer cachexia.

Impaired in vivo tumor growth of human pancreatic carcinoma cells retrovirally transduced with GM-CSF gene.

Year 1998
Kimura M. Tagawa M. Yoshida Y. Takenouchi T. Takenaga K. Azuma K. Yamaguchi T. Saisho H. Sakiyama S.
Division of Pathology, Chiba Cancer Center Research Institute, Japan.
We have examined the antitumor effect of human pancreatic carcinoma cells (AsPC-1) retrovirally transduced with mouse granulocyte macrophage-colony stimulating factor (GM-CSF) gene in nude mice. Growth retardation of the subcutaneous tumors of GM-CSF-producing AsPC-1 cells was observed, although their in vitro proliferation was not different from that of wild-type cells. Histological examination revealed infiltration of monocytic cells into the tumor of GM-CSF-producing cells, and they were shown to be mainly CD11b positive cells by immunohistochemical staining. The survival of the mice inoculated intraperitoneally with GM-CSF- producing AsPC-1 cells was significantly prolonged compared with that of the mice inoculated with wild-type AsPC-1 cells. Thus, the expression of GM-CSF gene in human pancreatic cells induced an antitumor effect in vivo even in the mature T cell-deficient condition.

Multiple synchronous colorectal carcinomas: a ploidy study by image analysis.

Year 1998
Caruso ML. Armentano R.
Servizio di Anatomia Patologica, I.R.C.C.S. S.de Bellis, Castellana Grotte, Bari, Italia.
Cytometry could represent an ancillary technique to morphology in order to understand whether multiple synchronous colorectal carcinomas arose independently. Twenty-eight multiple synchronous tumours, assessed by means of a computerized image analysis system for DNA ploidy, were categorized as diploid (4) or not diploid (24). The ploidy classes were: DNA-diploid, DNA-tetraploid, and DNA-aneuploid. The DNA Index (DI) ranged from 0,90 to 2,66. The overall concordance rate for ploidy and DI class with synchronous tumours was 69% and 31%, respectively. The high concordance rate in ploidic categories suggests the metastatic origin of our multiple synchronous tumours. Moreover, they showed site, Dukes' classification, degree of differentiation, percentage of ploidy class, and DI distribution comparable to the single colorectal carcinomas. In conclusion, image analysis is a reliable technique to determine the independent clonality or the common origin of multiple colorectal tumours when the evaluation based on the simple histopathological criteria is not satisfactory.

Clinicopathologic differences between gastric remnant cancer and primary cancer in the upper third of the stomach.

Year 1998
Imada T. Rino Y. Takahashi M. Shiozawa M. Hatori S. Noguchi Y. Amano T. Kobayashi O. Sairenji M. Motohashi H.
First Department of Surgery, Yokohama City University, School of Medicine, Japan.
BACKGROUND: The prognosis of gastric remnant cancer is considered to be poor compared with that of primary cancer. MATERIALS AND METHODS: This study was conducted to elucidate the differences of clinicopathologic findings and treatment results between 59 patients with gastric remnant cancer (GRC) and 579 patients with primary proximal gastric cancer (PGC). RESULTS: There was no significant difference in the incidence of lymph node metastasis between GRC and PGC. However, changes in the metastatic pattern to lymph nodes were observed in GRC. In GRC, the tumor easily invaded the neighboring organs due to the adhesions around the remnant stomach, resulting in a low resectability with curative intent. The 5-year survival rate after curative resection for advanced GRC was 50.9%. GRC patients without serosal invasion had a good prognosis. CONCLUSION: Although the survival rate after curative resection for GRC patients was similar to that of PGC, GRC patients without serosal invasion had a better prognosis. Therefore, early detection is an important way to improve overall survival in GRC.

Establishment of a liver metastatic model of human ovarian cancer.

Year 1998
Yoshida Y. Kamitani N. Sasaki H. Kusumi K. Tominaga T. Kotsuji F.
Department of Obstetrics and Gynecology, Fukui Medical School, Japan.
We have established an in vivo experimental model in which human ovarian cancer grows in the ovary of nude mice and metastasizes to parenchymatous organs. An ovarian cancer cell line was orthotopically injected into the nude mouse ovary together with Matrigel by microsurgical techniques. The cells grew locally in the ovary and metastasized to the peritoneum, colon, omentum, liver, and spleen. When the cells were injected into the intraperitoneal cavity with Matrigel, they formed carcinomatous peritonitis but neither ovarian tumor formation nor the metastasis to parenchymatous organs was detected. Taken together, these findings indicate that the microenvironment of the ovary seems to be essential for metastasis of implanted human ovarian cancer cells. This in vivo experimental model allows us to investigate the mechanism of the metastasis of ovarian cancer, it will be also useful for the establishing a new therapeutic approach to preventing metastasis of human ovarian cancer to parenchymatous organs.

Antitumor effect of human pancreatic cancer cells transduced with cytokine genes which activate Th1 helper T cells.

Year 1998
Yoshida Y. Tasaki K. Kimurai M. Takenaga K. Yamamoto H. Yamaguchi T. Saisho H. Sakiyama S. Tagawa M.
Division of Pathology, Chiba Cancer Center Research Institute, Japan.
We have examined antitumor effect of human pancreatic carcinoma cells (AsPC-1) retrovirally transduced with interleukin-12 (IL-12), IL-15 or IL-18 gene in nude mice. The tumor growth of IL-12-expressing AsPC-1 cells was significantly retarded and that of IL-15-expressing cells was also impeded compared with that of wild-type cells, although their in vitro cell growth remained unchanged. However, the expression of IL-18 in AsPC-1 cells did not generate any antitumor effect since the tumor growth of the transduced cells was the same as that of wild-type cells. Thus, the differential actions of these cytokines on non-T cells can generate a variety of antitumor effect in nude mice, although their actions on T cells lineage favor the stimulation of Th1-type helper T cells.

Enhancement of anticancer effects of radiation and conventional anticancer agents by a quinolinone derivative, vesnarinone: studies on human gastric cancer tissue xenografts in nude mice.

Year 1998
Kawai K. Konishi Y. Izumi K. Sato M. Adachi M. Hozumi M.
Cellular Technology Institute, Otsuka Pharmaceutical Company, Tokushima, Japan.
Vesnarinone (3,4-dihydro-6-[4-(3,4-dimethoxybenzoyl)-1-piperazinyl]-2(1H)- quinolinone), a quinolinone derivative, is an orally active inotropic agent used in Japan for the treatment of chronic heart failure. Recently, it has been reported that vesnarinone induces differentiation and apoptosis in certain types of leukaemia and solid tumour cells, and exhibits antitumour effect on several tumours xenografted in nude mice. In the present study, we examined the antitumour effect of vesnarinone in combination with radiation and conventional anticancer agents in nude mice xenografted with human gastric carcinoma, a poorly-differentiated adenocarcinoma, MKN-45 cell line which has a wild-type p53 gene. Vesnarinone treatment combined with radiation resulted in a higher antitumour activity compared with a single treatment with either vesnarinone or radiation alone. Further, vesnarinone treatment together with radiation and conventional anticancer agents including 5-FU and picibanil (an immunopotentiator) produced the highest antitumour effect compared with any other treatment. Additionally, the combination treatment induced marked differentiation and apoptosis of the tumour cells and an increase in the expression of p53 gene in the treated tumour cells. The results suggest that vesnarinone, in combination with radiation and the conventional antitumour agents, may be of clinical interest for treatment of certain types of gastric tumours.

beta-Sitosterol inhibits growth of HT-29 human colon cancer cells by activating the sphingomyelin cycle.

Year 1998
Awad AB. von Holtz RL. Cone JP. Fink CS. Chen YC.
Department of Physical Therapy, Exercise and Nutrition Sciences, SUNY/Buffalo, NY 14214, USA. awad@acsu.buffalo.edu
The present study examined the SM cycle as a mechanism to explain the inhibitory effect of SIT on HT-29 cell growth. SIT was the main phytosterol in the diet. Supplementation of SIT at 16 microM for 5 days in the media inhibited growth by 55% as compared to cholesterol. SIT supplementation had no effect on sphingosine production. Ceramide production increased 45% with SIT supplementation as compared to cholesterol. Sterol supplementation had no effect on phospholipase C, a key enzyme in the PKC pathway. We concluded that the activation of the SM cycle may play a role in growth inhibition of HT-29 cells by SIT.

Radiochemosensitivity and expression of p53 in patients with esophageal cancer treated by absolute noncurative resection.

Year 1998
Ikeguchi M. Saito H. Katano K. Kondou A. Tsujitani S. Maeta M. Kaibara N.
Department of Surgery I, Faculty of Medicine, Tottori University, Yonago, Japan.
The aim of this study was to investigate the relationship between the expression of p53 protein and radiochemosensitivity in patients with advanced esophageal cancer. We analyzed the expression of p53 protein by an immunohistochemical method in tumors from 28 patients with advanced esophageal cancer who had been treated by absolute noncurative resection. Four patients died from postoperative complications. Fourteen of the remaining 24 patients were treated with postoperative radiochemotherapy. The expression of p53 protein was detected in tumors from 16 of 28 patients (57.1%). The mean survival period was 16.4 months for 14 patients treated with radiochemotherapy, as compared to 6.9 months for 10 patients who were not treated with radiation therapy (P = 0.112). In the case of the 14 patients treated with radiochemotherapy, the mean survival period was 24.7 months for seven patients with p53-negative tumors while it was only 8 months for seven patients with p53-positive tumors. However, difference was not statistically significant (P = 0.149). Postoperative radiochemotherapy prolonged the survival of some patients with noncuratively operated esophageal tumors that were p53-negative. However, the enhanced survival was not statistically significant.

Activity of high dose 24 hour 5-fluorouracil infusion plus L-leucovorin in advanced colorectal cancer.

Year 1998
Nobile MT. Barzacch MC. Sanguineti O. Chiara S. Gozza A. Vincenti M. Lavarello A. Cognein P. Lionetto R. Percivale PL. Bertoglio S. Murolo C. Esposito M. Vannozzi MO. Rosso R.
Istituto Nazionale per la Ricerca sul Cancro, Genova, Italy.
BACKGROUND: Modulation of 5-fluorouracil (5-FU) by leucovorin (L-LV) in patients (pts) with advanced colorectal cancer has been demonstrated to produce a highly significant benefit over single-agent 5-FU in terms of tumor response rate, but this advantage does not translate into an evident improvement of overall survival. To improve the clinical efficacy of the 5-FU plus L-LV regimen a phase II study of weekly 24-hour high-dose 5-FU infusion with L-LV was undertaken. PATIENTS AND METHODS: Seventy advanced colorectal patients were enrolled and treated by a weekly outpatient combination regimen according to the following schedule: L-LV 100 mg/sqm by 4 hours i.v. infusion followed by 5-FU 2600 mg/sqm over a 24 hours infusion combined with a fixed dose of oral L-LV (50 mg) every 4 hours for 5 times. Forty-four pts did not receive any previous CT and 26 pts were pretreated with fluoropyrimidines. RESULTS: The overall objective response rate (OR) was 35.3%; 7 CR and 11 PR (42.8% OR) were observed in the group of untreated pts, and 6 PR (23% OR) were reported among previously treated pts. Major side effects were represented by diarrhoea (grade III: 26%, grade IV: 1%), hand-foot syndrome (grade III: 4%, grade IV: 1%) and mucositis (grade III: 4%); however, this did not significantly influence the therapeutic programme. Median 5-FU dose intensity was 100% and 80% at 4 weeks, 87% and 75% at 8 weeks in untreated and pretreated pts, respectively. CONCLUSIONS: L-Leucovorin modulation of weekly short-term continuous infusion of high-dose 5-fluorouracil appeared a well-tolerated outpatient regimen; it demonstrated a high activity in advanced colorectal cancer, both in untreated pts and in pts resistant to 5-FU-based chemotherapy.

Neoadjuvant chemotherapy of gastric cancer with oral UFT (a mixture of uracil and fturafur) during the waiting period for surgery.

Year 1998
Nio Y. Sato Y. Nagami H. Teramoto M. Inoue Y. Yano S. Sumi S. Tamura K. Fukumoto M.
First Department of Surgery, Shimane Medical University, Shimane, Japan.
Our previous experience has demonstrated that growth of gastric cancer during the waiting period for surgery cannot be neglected, and some patients hope to receive prophylactic treatment to inhibit the growth of tumor until surgery. The present study was designed to assess the clinical benefits of preoperative chemotherapy with oral UFT for gastric cancer during the waiting period for surgery. Fifty patients with gastric cancer (24 early, 25 advanced and 1 recurrent cancers) were treated with oral UFT at 300-600 mg/day for 7-36 days before surgery and the objective responses and the postsurgical survivals were evaluated. In 42 of 50 patients objective responses of primary lesions were assessed by endoscopy or upper gastrointestinal series examination, and 2 CRs, 15 PRs and 25 NCs were seen (40% response). The histological effect was evaluated in 50 patients and the following classifications were made: grade 3 (complete disappearance or necrosis of tumor cells), 2; grade 2 (necrotic changes > 2/3 area), 4; grade 1b (> 1/3 area), 7; grade 1a (< 1/3 area), 15; and grade 0 (no histological changes), 22. A longer period of UFT administration was associated with CR or PR. All the patients underwent gastrectomy (38 curative and 12 palliative gastrectomies): all patients with Stage I-III primary gastric cancer are alive after surgery, and the 50% survival period of the patients with Stage IV cancer was 20 months. The side effects were not serious, including slight myelotoxicity, liver dysfunction and anorexia. It is concluded that preoperative chemotherapy for gastric cancer with oral UFT on outpatient basis may result in down-staging as well as the prevention of tumor growth during the waiting period for surgery without serious side effects.

Significance of nm23 mRNA expression in human hepatocellular carcinoma.

Year 1998
Lin LI. Lee PH. Wu CM. Lin JK.
School of Medical Technology, College of Medicine, National Taiwan University, Taipei, Taiwan.
It has been proposed that nm23-H1, a candidate suppressor gene for metastasis, plays an important role in the metastasis of human tumors. In order to investigate its role in the human hepatocellular carcinoma (HCC), 18 matched pairs of tumorous and adjacent nontumorous liver tissues of hepatectomy from patients with HCC were studied by quantitative reverse transcription-PCR. Further analyses of the nm23-H1 gene were also done. The results of these molecular studies were correlated with the clinicopathologic features of the patients. Nm23-H1 transcript was expressed in all neoplastic and adjacent nontumorous liver tissue. The level of expression, however, did not correlate well with the extension or metastatic potential of the tumors. Instead, in 15 (83.3%) of 18 HCC, nm23-H1 expression was higher in the tumorous tissues, compared with the adjacent nontumorous tissues; and significantly higher levels of nm23-H1 mRNA expression was detected in HCC with poor differentiation (Edmonson classification, III and IV) than those with moderate differentiation (I and II). Southern blot analysis of nm23-H1 gene revealed neither amplification nor loss of heterozygosity of all HCC tissues examined. Direct sequencing of the nm23-H1 gene in all HCC tissues detected no mutations. Our findings suggested that increased nm23-H1 mRNA expression is correlated with HCC tumor progression.

Protein expression of p53, p21WAF1, and Rb as prognostic indicators in patients with surgically treated hepatocellular carcinoma.

Year 1998
Naka T. Toyota N. Kaneko T. Kaibara N.
First Department of Surgery, Faculty of Medicine, Tottori University, Japan.
Many genes participate in the regulation of cell cycle progression from G1 to S phase. Functional loss of one or more of these genes has been reported to be associated with carcinogenesis and/or tumor progression and poor prognosis in many cancers. In a series of 126 patients with hepatocellular carcinoma (HCC), we immunohistochemically evaluated tumor expression of the cell cycle-related gene protein products of Rb, p21 (WAF1), and p53. Positive immunostaining for Rb, p21, and mutant p53 protein was detected in 58%, 33%, and 37% of the tumors, respectively. The proportion of HCCs exhibiting aberrant p53 protein expression increased significantly with advancing stage of disease (p < 0.001), poorer histological classification of differentiation (p < 0.01), and increasing tumor size (p < 0.01). A decrease in the proportion of HCCs expressing p21 protein was also associated with advancing clinical stage of disease (p < 0.01), and larger tumor size (p < 0.05). The only clinicopathological feature found to be associated with Rb status, was intrahepatic metastasis, which occurred with a higher frequency in HCCs exhibiting positive immunoreactivity for Rb protein expression (p < 0.05). Multivariate survival analysis revealed that, amongst the protein products of the different genes evaluated, only positive immunostaining for aberrant p53 protein expression served as an independent prognostic indicator, being significantly associated with worse survival in patients with HCC (p = 0.023). Analysis for relationships between gene products showed an inverse correlation between expression of aberrant p53 protein and p21 protein (p < 0.01), and also an inverse correlation between p21 protein and Rb protein expression (p < 0.05) in these cases of HCC. These findings demonstrate that positive immunostaining for mutant p53 protein expression is a significant indicator of tumor progression and poor prognosis, confirm that p21 protein expression is induced in a p53-dependent manner, and suggest that Rb protein expression may be regulated to some extent by p21 in HCC.

Clinical contribution of bcl-2, p53 and Ki-67 proteins in pancreatic ductal adenocarcinoma.

Year 1998
Makinen K. Hakala T. Lipponen P. Alhava E. Eskelinen M.
Department of Surgery, University Hospital and University of Kuopio, Finland.
The expression of apoptosis suppressing protein bcl-2, tumour-suppressor protein p53, and proliferation marker Ki-67 and their possible prognostic value were analysed in pancreatic ductal adenocarcinoma. Fifty-two % (34/64) of the samples were positive for bcl-2 and immunostaining were mainly localized in the cytoplasm of tumour cells. Bcl-2 expression was not related to tumour grade, DNA ploidy or S-phase fraction or to any clinical parameters. In univariate analysis bcl-2 expression predicted favourable outcome (p = 0.008). Positive nuclear staining for p53 was found in 40% (24/59) of samples and 80% (60/74) of the tumours were positive for Ki-67. p53 and Ki-67 expressions were not related to patient survival. According to our results, bcl-2 expression seems to be a predictor of disease outcome and may have some clinical value in human pancreatic cancer.

Five-lipoxygenase inhibitors reduce Panc-1 survival: the mode of cell death and synergism of MK886 with gamma linolenic acid.

Year 1998
Anderson KM. Seed T. Meng J. Ou D. Alrefai WA. Harris JE.
Department of Medicine, Rush Medical College, Chicago, IL 60612, USA. Kanderso@Rush.Edu
The 5-lipoxygenase inhibitors ETYA, SC41661A and MK886 reduced the proliferation and viability of Panc-1 human pancreatic cancer cells. The extent of inhibition depended upon drug concentration, and with continued culture, cells detached and stained with trypan blue. Although results from flow cytometry were those associated with programmed cell death, despite repeated attempts, no DNA laddering consistent with its later stages was detected, and studies with the TUNEL assay were negative. Light and electron microscopy of cells cultured with SC41661A provided morphologic evidence of a population of "dark" cells and of an incompletely expressed type 1 programmed cell death including margination of chromatin at the nuclear membrane and by consolidation and degeneration of cytoplasmic organelles, along with extensive vacuolization. Cells cultured with MK886 exhibited compact "dark" cells and an unusual cytoplasmic mode of cell death characterized by vacuolization and widely separated smooth internal membranes without diagnostic nuclear changes. This is in marked contrast to the extensive type 1 PCD induced by 5-lipoxygenase inhibitors cultured with human U937 monoblastoid cells. On balance, the response of Panc-1 cells to MK886 suggests expression of a variant type 2 (autophagic) cellular suicide, although some contribution from components of a "cytoplasmic" (type 3?) form of non-necrotic cell death may also be considered. In a European clinical trial, gamma linolenic acid, a polyunsaturated fatty acid that generates free radicals has been combined with 5-fluorouracil as chemotherapy for pancreatic cancer. Panc-1 cell proliferation was insensitive to inhibition by several chemotherapeutic agents employed clinically, including 5-fluorouracil, cisplatin or gemcitabine and only somewhat sensitive to GLA. When gamma linolenic acid was combined with MK886, the more effective of the two 5-lipoxygenase inhibitors, a synergistic reduction in Panc-1 cell number and viability occurred.

Adjuvant 131I-anti-CEA-antibody radioimmunotherapy inhibits the development of experimental colonic carcinoma liver metastases.

Year 1998
Mahteme H. Lovqvist A. Graf W. Lundqvist H. Carlsson J. Sundin A.
Department of Surgery, Uppsala University, Sweden.
Adjuvant radioimmunotherapy (RIT) for human colonic cancer was performed in a nude rat model of experimental liver metastases. Thirty-three rats were injected intraportally through a mesenteric vein with 5 x 10(6) cells from the human colonic cancer cell line LS174T. Within half an hour, 20 MBq (n = 2), 75 MBq (n = 5), or 150 MBq (n = 10) of the 131I-labelled anti- carcinoembryonic antigen (CEA) monoclonal antibody (MAb) 38S1 was administered intravenously (i.v.), whereas control groups received either i.v. saline injections (n = 12) or 150 MBq of the irrelevant 131I-labelled MAb 79C (n = 4). Decay corrected whole-body data showed that more than 80% of the initially MAb-bound radioiodine was excreted during the first 2 weeks. Whole- body clearance and blood clearance of 131I-38S1 and 131I-79C were essentially similar. At sacrifice 5-7 weeks after administration, neither 20 MBq nor 75MBq 131I-38S1 significantly prevented the development of liver metastases. By contrast, with 150 MBq, no metastases formed in the animals treated with MAb 131I-38S1 or 131I-79C. A radiation induced effect on the haematopoietic system was found in the 150MBq dosage groups. It is concluded that the inhibition of tumour induction was not strictly dependent on a radiation dose delivered by a tumour-specific MAb. Since a non-tumour-specific 131I-MAb, in a smaller group of animals, proved equally efficacious in preventing tumour growth, the total body 131I dose was probably the major contributing factor.

Vascular endothelial growth factor expression is reduced in liver metastasis from colorectal cancer and correlates with urokinase-type plasminogen activator.

Year 1998
Berney CR. Yang JL. Fisher RJ. Russell PJ. Crowe PJ.
Department of Surgery, University of New South Wales, Randwick, Australia.
We undertook a retrospective study to investigate the correlation between the expression of vascular endothelial growth factor (VEGF) and urokinase-type plasminogen activator (u-PA) proteins with progression of colorectal carcinoma (CRC). Immunohistochemical analyses using antibodies against VEGF and u-PA were carried out on archival specimens of 58 human colon carcinomas, 30 liver secondaries and 20 adenomas. Expression of VEGF was significantly reduced in the metastatic liver tumours compared with primary ones (Wilcoxon test, P = 0.002), suggesting VEGF activity to be secondarily down-regulated once the tumour cells reach the hepatic parenchyma. There was no strong evidence from our data that the level of VEGF or u-PA in the primary tumour could predict risk of liver metastasis or survival duration. VEGF and u-PA expression were positively correlated in primary CRC suggesting that both proteins may interact in vivo (chi-square test, P = 0.019) in tumour progression.

Genetic instability, p53 and nm23 mutation and clinicopathological features in rectal carcinoma.

Year 1998
Indinnimeo M. Cicchini C. Stazi A. Mingazzini P. Caligo MA. Ghimenti C. Bevilacqua G.
First Department of Surgery, University of Rome La Sapienza, Rome, Italy.
Defective DNA mismatch repair proteins fail to correct replication errors (RERs). These defects may lead to secondary, mutation of oncogenes and tumor suppressor genes. Microsatellite instability might be a marker of such replication errors. Eighteen rectal tumors were examined to evaluate genetic instability, in sporadic rectal cancer by PCR. RERs were observed in 27.8% of the cases. No significant difference was noticed between RER+ and RER- patients as far as prognosis, clinicopathological features and p53 gene mutation are concerned. The incidence of nm23 gene mutation was the only statistically significant difference between the 2 groups. Three patients with only one altered microsatellite showed advanced tumor and nm23 gene mutation. Two cases with 5 altered microsatellites and nm23 gene mutated are disease-free: in one of them the p53 gene was also mutated. Probably more than one altered microsatellite is necessary to protect from the effects of secondary mutations.

Chemosensitivity of human pancreatic cancer cell lines serially transplanted in nude mouse.

Year 1998
Tomikawa M. Kubota T. Takahashi S. Matsuzaki SW. Kitajima M.
Department of Surgery, School of Medicine, Keio University, Tokyo, Japan.
BACKGROUND: Pancreatic cancer frequently recurs or metastasizes even after apparently curative surgical resection. Because of a low, five-year survival rate after radical surgery, multi-modal adjuvant treatment must be used to prevent recurrence of systemic spread. MATERIALS AND METHODS: The effectiveness of the experimental cancer chemotherapy of mitomycin C (MMC), cisplatin (DDP), doxorubicin (DXR) and 5-fluorouracil (5-FU) was evaluated in three human pancreatic cancer xenografts serially transplanted in nude mice. RESULTS: When the effects of these agents were evaluated by 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay, only MMC and DDP were effective on PAN-3-JCK, a poorly differentiated adenocarcinoma. When PAN-12-JCK, a moderately differentiated adenocarcinoma, was used an in vitro assessment of combined chemotherapy of MMC and DDP, a synergistic combination effect was observed. Three xenografts were transplanted subcutaneously into nude mice and the maximum tolerated doses of these agents were administered intraperitoneally or intravenously (DXR). MMC showed positive antitumor activity on PAN-3-JCK and PAN-12-JCK, and 5-FU was effective on PAN-12-JCK. CONCLUSIONS: These results reflect the low sensitivity of clinical pancreatic cancer to conventionally available antitumor agents, and suggest the possible synergistic combination antitumor activity of MMC and DDP.

Alkaline phosphatase isoenzymes in detection and follow up of breast cancer metastases.

Year 1998
Ritzke C. Stieber P. Untch M. Nagel D. Eiermann W. Fateh-Moghadam A.
Institute of Clinical Chemistry, Ludwig-Maximilians-University Munich, Germany.
Alkaline phosphatase isoenzymes in bone and liver metastases of breast cancer were determined in 637 patients and during follow up in 116 patients, by agarose gel electrophoresis. The activity of total alkaline phosphatase, gamma glutamyl transferase, CEA and CA 15-3 was also determined. The sensitivity of bone alkaline phosphatase in bone metastases detection was 39%, of electrophoresis (total alkaline phosphatase and specific isoenzymes) 48% and of enzymes and tumor markers together 85% (n = 62). For liver metastases gamma glutamyl transferase had the best sensitivity (100%, n = 19). Even though positive predictive value was only 6%, the sensitivity of liver alkaline phosphatase was 58% and of electrophoresis 94%. The determination of alkaline phosphatase isoenzymes is a non-invasive, in-expensive, reproductable and rapid method to detect progressive disease in breast cancer, especially in combination with the tumor markers CA 15-3 and CEA.

Cisplatin, etoposide, and weekly high-dose 5-fluorouracil and leucovorin infusion (PE-HDFL)--a very effective regimen with good patients compliance for advanced gastric cancer.

Year 1998
Cheng AL. Yeh KH. Lin JT. Hsu C. Liu MY.
Department of Internal Medicine, National Taiwan University Hospital, Taipei.
We have previously shown that weekly 24-hour infusion of high-dose 5-fluorouracil and leucovorin (HDFL), a regimen initially designed for the treatment of advanced colorectal cancer, is also effective in the treatment of gastric cancer. This HDFL regimen is unique in that it is virtually non-myelosuppressive, and thus provides a comerstone on which ideal protocols may be developed. In this prospective phase II study, we examined the efficacy and toxicity of PE (cisplatin, etoposide)-HDFL, a HDFL-based combination chemotherapy, in the treatment of advanced-gastric cancer. This regimen consisted of cisplatin 60 mg/m2, i.v., D1; etoposide 65 mg/m2, i.v., D1-3; and 5-fluorouracil 2600 mg/m2 plus leucovorin 300 mg/m2, 24-hour i.v. infusion by an ambulatory infusion pump, D2,9,16; repeated every 4 weeks. The major eligibility criteria of the patients included: a) a histologically confirmed, objectively measurable, recurrent or primary inoperable gastric adenocarcinoma; b) age > or = 75 years; c) a Karnofsky performance status > or = 50%; d) an absolute granulocyte count (AGC) > or = 2000/mm3, and a platelet count > or = 100,000/mm3; e) a serum bilirubin concentration < or = 2.0 mg/dl; f) a serum creatinine concentration < or = 1.5 mg/dl; and g) a signed informed consent. Between March 1992 and June 1996, a total of 42 patients were enrolled onto the study. There were 31 men and 11 women with a median age of 54 (24-75) years; these included 16 primary metastatic, 3 locally advanced and inoperable, and 23 postgastrectomy recurrent gastric cancer patients. ECOG (Eastern Cooperative Oncology Group) grade III/IV leukopenia and thrombocytopenia developed in 34.0% and 11.0% of a total of 229 courses given, respectively. There was no treatment-related death. Four patients developed a reversible neurotoxicity; and two of them refused further chemotherapy. Among the 40 patients evaluable for responses, 9 [22.5%; 12-38%, 95% confidence interval (C.I.)] patients achieved complete remission, and 20 [50.0%; 33-67%, 95% C.I.] patients achieved partial remission. The overall response rate was 72.5% [56-86%, 95% C.I.]. The overall median survival and median time to progression of the responders were 10 and 7 months, respectively. The overall median survival of the whole group was 9 months. We concluded that PE-HDFL is a highly effective treatment for advanced gastric cancer. The treatment-related toxicity was mild and the patients' compliance was satisfactory.

Preliminary report of a controlled trial of MTH-68/B virus vaccine treatment in acute B and C hepatitis: a phase II study.

Year 1998
Csatary LK. Telegdy L. Gergely P. Bodey B. Bakacs T.
United Cancer Research Institute, Alexandria, VA, USA.
Eighty four patients with viral hepatitis attributed to infection with hepatitis B virus (HBV) (n = 43) or hepatitis C virus (HCV) (n = 41) were included in this study employing the MTH-68/B vaccine, an attenuated variant of Bursal Disease Virus. Twenty of the 43 patients in the HBV group, and 22 of the 41 HCV patients were treated with MTH-68/B. The remaining patients received conventional therapy. Significantly more patients progressed into active chronic hepatitis on conventional therapy (13% of HBV and 26% of HCV cases respectively) than in the vaccine treated groups (0% and 9%). Relapses occurred less frequently in the vaccine treated groups (5% of HBV and 32% of HCV) than in the control groups (9% and 79%), while remissions within one month of treatment were observed more often in the vaccine treated groups (both 50% respectively) than in the control groups (26% of HBV and 21% of HCV patients respectively). The duration of hepatitis was also considerably shortened by MTH-68/B treatment in both HBV (from 7.5 +/- 3.7 to 5.9 +/- 3.0 weeks) and HCV patient groups (from 8.9 +/- 7.4 to 5.3 +/- 4.4 weeks). The data presented suggest that attenuated, non-pathogenic viruses may be of significant benefit for patients with viral hepatitis B and C infections.

Detection of circulating cancer cells by nested reverse transcription-polymerase chain reaction of cytokeratin-19 (K19)--possible clinical significance in advanced gastric cancer.

Year 1998
Yeh KH. Chen YC. Yeh SH. Chen CP. Lin JT. Cheng AL.
Department of Oncology, National Taiwan University Hospital, Taipei, R.O.C.
Intermediate filament cytokeratin-19 (K19) protein is expressed in normal and malignant gastrointestinal epithelial cells, but not in peripheral blood (PB). Small amount of circulating gastric cancer cells can be detected by a sensitive nested reverse transcription-polymerase chain reaction (RTPCR) with primers specific for K19 mRNA. Thirty-four PB samples obtained from patients with inoperable/metastatic gastric cancer were examined. The mononuclear cell (MNC) fraction was collected by Ficoll centrifugation, and followed by total RNA extraction by acid guanidinium thiocyanate-phenol-chloroform method. RNA from 8 gastric cancer cell lines and the mononuclear cells of 33 healthy adults were used as positive and negative controls, respectively. DNA fragment of 774 bp amplified by the internal primers was found to be a highly reliable marker for K19 mRNA expression. The sensitivity of detection was between 1 and 10 cells/10(6) normal MNCs. The K19 transcripts were detected in 20.6% (7/34; 8-37%, 95% C.I.) of PB samples. None of the other pertinent clinicopathological features, including the disease extent and the histopathologic types of the tumors, were related to the expression of K19 in PB. All 34 patients had been treated by systemic chemotherapy. Among the 17 non-responders to chemotherapy, the survival of the 4 patients with detectable K19 was significantly shorter than that of 13 patients without detectable K19 in their circulating blood (p = 0.014). However, the survival impact of K19 was less significant in the other 17 patients whose tumors had responded to systemic chemotherapy. Of the whole group of patients, the median survival of the 7 and 27 patients with and without detectable K19 in their circulating blood was 1 and 3.5 months, respectively (p = 0.368). We concluded that detecting circulating cancer cells by K19 nested RT-PCR is associated with poor prognosis of gastric cancer, particularly in those patients who are not responsive to systemic chemotherapy.

Elevated reversible and irreversible lipid peroxidation in human oesophageal cancer.

Year 1998
Levy RD. Oosthuizen MM. Degiannis E. Lambrechts H.
Department of Surgery, University of the Witwatersrand Medical School, Johannesburg, South Africa.
BACKGROUND: Cancer of the oesophagus is the most common gastrointestinal malignancy in South African blacks. The aim of this study was to determine the reversible and irreversible lipid peroxidation in cancer of the oesophagus; a feature that has not been assessed before in this or other cancer tissue. METHODS: Biopsies and plasma from 39 patients with cancer of the oesophagus and 22 biopsies and plasma from non-cancer patients were analysed for the irreversible lipid peroxide product malondialdehyde (MDA) and other reversible lipid peroxide products (LPO) by the thiobarbituric acid-reactive substances (TBARS) method. RESULTS: The mean (+/- SEM) for MDA in plasma from normal patients was 1.697 (0.149), and for cancer patients 4.23 (0.417) nmol MDA/ml. The tissue MDA for normal patients was 0.807 (0.154), and for cancer patients 2.530 (0.379) nmol MDA/mg protein. The mean (+/- SEM) for LPO in plasma from normal patients was 1.929 (0.281), and for cancer patients 12.607 (1.451) nmol MDA/ml. The tissue LPO for normal patients was 2.957 (0.306), and for cancer patients 16.320 (1.868) nmol MDA/mg protein. All the MDA and LPO values for cancer patients were significantly elevated (p < 10(-4)). In oesophageal cancer 85% of the lipid which was peroxidized, was of the reversible type. CONCLUSIONS: This elevated reversible LPO levels in plasma and oesophageal tissue of cancer patients support the notion that the oxy-radicals in cancer are not under proper metabolic control. Therefore, human oesophageal cancer does not progress to self regression or destruction but rather to more mutagenic changes probably due to high reversible lipid peroxidations.

Ethnic or environmental differences in disparate geographic regions may influence the histology of flat colorectal neoplasias.

Year 1998
Rubio CA. Saito T. Kawaguchi M. Miyaoka M. Horimukai H. Taguchi Y. Katayama A. Jaramillo E. Watanabe M. Koizumi K. Slezak P. Karu S. Hirota T.
Department of Pathology, Karolinska Institute and Hospital, Stockholm, Sweden. Carlos.Rubio@onkpat.ki.se
Histologic sections of endoscopically flat colorectal polyps removed in Tokyo and Stockholm were reviewed. A total of 178 flat colorectal neoplasias (88 from the Tokyo Medical College Hospital, Tokyo and 90 from the Karolinska Hospital, Stockholm) were classified following strict histologic criteria by two different pathologists (one Swedish and the other Japanese). The number of polyps with high grade dysplasia, with intramucosal carcinoma and with invasive carcinoma were significantly higher (p < 0.001) in Tokyo (61.4% or 54/88) than in Stockholm (15.0% or 14/90). The present results suggest that flat colorectal neoplasic polyps may be histologically more "severe" and more "aggressive" in Japanese than in Swedish patients. The possibility that more "advanced" lesions had been inadvertently removed in Tokyo was discounted as Japanese endoscopists were also instrumental in excising many of the flat colorectal polyps in Stockholm. Ethnic and/or environmental differences seem to play a crucial role in the evolution of flat colorectal neoplasic polyps from LGD and HGD, to intramucosal and to invasive carcinoma.

Histopathologic prognostic score in colorectal adenocarcinomas.

Year 1998
Diez M. Pollan M. Enriquez JM. Dominguez P. Santana A. Tobaruela E. Muguerza JM. Arrieta F. Rodriguez A. Ruiz A.
Hospital Principe de Asturias, Madrid, Spain.
OBJECTIVE: To analyze the prognostic value of a set of pathological variables after curative resection for large bowel adenocarcinoma and to test a prognostic score derived from factors with independent effect. PATIENTS AND METHODS: The study is based on data from 292 consecutive unselected patients (B-C Astler-Coller stages). Histopathological features were evaluated prospectively on the resected primary tumors. Relationship between these factors and risk of recurrence was assessed by a Cox's proportional regression analysis. RESULTS: Four variables retained independent prognostic significance: extent of bowel wall invasion, peritumoral lymphocytic infiltration, number of positive nodes and vascular invasion. A prognostic score based on the regression coefficients attained by such variables was developed. This system revealed four prognostic groups. Group I included 14% of patients, with 94% 5-year disease-free survival. These figures were: 35% and 60% in group II; 43% and 46% in group III; and 7% and 24.4% in group IV. Histopathologic score applied to bearers of Astler-Coller B2 tumors permitted the identification of two populations, one characterized by a low risk of relapse and another with high risk (p = 0.002). CONCLUSION: A prognostic score based in the evaluation of four histopathologic parameters concerning the tumor phenotype enables the identification of groups of patients at risk of relapse after curative resection for colorectal adenocarcinoma.