beta-Lapachone induced cell death in human hepatoma (HepA2) cells.
Lai CC. Liu TJ. Ho LK. Don MJ. Chau YP.
Institute of Anatomy, School of Life Science, National Yang-Ming University, Taipei, Taiwan, Republic of China.
In present study we studied the cytotoxic effects of beta-lapachone, a potent anticancer drug, on the human hepatoma cell line (HepA2) under serum-free condition. Most cells died after 2 microM beta-lapachone addition at 48 hours. No apoptotic characteristics of DNA ladder was documented by agarose DNA electrophoresis. The blockage of cell cycle at S phase and unscheduled DNA synthesis were demonstrated by flow cytometric analysis and anti-bromodeoxyuridine immunocytochemistry. Ultrastructural observation showed that the swollen mitochondria, dilatation and vesiculation of rER and proliferation of peroxisome-like granules appeared within the cytoplasm of HepA2 cells following drug treatment. Using enzyme cytochemistry, both peroxidase and acid phosphatase activities but not catalase activity were localised in these peroxisome-like granules. Therefore, these results suggested that (a) beta-lapachone has a novel cytotoxic effect on human hepatoma cell; (2) beta-lapachone induces the interruption of the cell cycle and unscheduled DNA synthesis in HepA2 cells; and (3) beta-lapachone promotes the proliferation of peroxisome-like granules containing peroxidase and acid phosphatase activities without evidence of catalase activity in hepatoma cell line.
An electron microscopic study of Helicobacter pylori in the surface mucous gel layer.
Ogata M. Araki K. Ogata T.
Department of Surgery, Kochi Medical School, Japan.
This study describes the distribution of spiral and coccoid forms of Helicobacter (H.) pylori within the surface mucous gel layer (SMGL) on human gastric mucosae. Gastric mucosae infected with H. pylori were obtained from surgically removed stomachs of 14 cases of gastric cancer. The glycocalyx of the spiral and coccoid forms of H. pylori was examined for specific lectin labeling. Coccoid forms were identified in 11 of these cases (78.6%). The SMGL, which was from 50 to 100 microns in thickness, contained H. pylori throughout its entire thickness. Variously-sized vacuole-like clear areas were present near H. pylori. The glycocalyx on both the spiral and coccoid forms was similar in its staining with the 8 types of lectins tested. However, the staining pattern of the lectins varied among different samples. The numerous fibrillae-like filaments radiated from the surface of the bacteria and appeared to link the bacteria to the surface mucous cells or the surrounding mucus. These fibrillae-like filaments were not specifically stained by the lectin reactions, suggesting absence of sugar molecules.
Quantitative assessment of normal and potentially premalignant epithelium at different levels of human colorectal crypts.
Tipoe GL. White FH.
Department of Anatomy, Faculty of Medicine, University of Hong Kong, Hong Kong.
The present study uses morphometric techniques to assess whether altered differentiation patterns exist in PPM which might reflect its premalignant status. Samples were obtained from resected malignant lesions of large bowels of 10 Chinese patients. Normal (N) samples were biopsied from the margins of each resected large bowel. Potentially premalignant (PPM) mucosae were obtained from within 2 cm of the margins of the malignant lesions. Tissues were processed for histological examination and using strict criteria, colorectal crypts were divided into basal (B), intermediate (I) and surface (S) segments. Interactive digitisation of sections from each group was used to generate the following morphometric parameters in each segment: nuclear profile circularity indices (NSF and NCI); nuclear numerical density (NA and NV); the degree of deviation of the major nuclear axis in relation to the epithelial-connective junction (AGDMAX); cell height (CH); the distance between nuclear apex to cell apex (DNACA); the distance between cell base to nuclear apex (DCBNA); stratification index (SI)--the ratio of DCBNA and CH; and the volume density of mucous vacuoles in the reference epithelium (VVMV,EP). In comparisons of different segments within groups, the nuclei at the S segment of N and PPM crypts were more irregular and less circular in shape than nuclei from other segments. There was a shift of nuclear profile shape (NSF and NCI) from circular to ellipsiodal between B and S segments. In comparisons of similar segments between groups, no significant nuclear shape changes were detected in nuclei of PPM crypts when compared with nuclei in similar segments of N crypts and the pattern of nuclear shape alterations resembled those of normal crypts. In comparisons of different segments within groups of N and PPM crypts, AGDMAX, DNACA, DCBNA, CH and SI parameters demonstrated that epithelial cells at the I segments have more centrally positioned nuclei with the tallest epithelial height when compared with epithelial cells in other segments of both crypts. In B segments, nuclear NA and NV were almost double those of other segments in both N and PPM crypts, with marked reductions in these parameters between B and I segments. VVMV,EP was significantly highest in the I segment and significantly lowest in the S segment of both groups. Both N and PPM crypts showed similar trends in VVMV,EP within the crypt segments but when comparing similar segments between both crypts, a significant difference was detected only between S segments. The alterations of nuclear shape and packing densities, orientation and mucous content in N crypts were similarly expressed in PPM crypts and distinct differences in numerical density (NV) and stratification index existed in crypts between these two groups when comparing similar segments. All values in PPM were consistently lower when compared with N crypts. These preliminary observations may represent a subtly altered state of cellular differentiation in PPM which may be a reflection of early preneoplastic transformation.
Chemopreventive effects of NSAIDs against colorectal cancer: regulation of apoptosis and mitosis by COX-1 and COX-2.
Department of Medicine, Hope Hospital, University of Manchester, Salford, UK.
There is a wealth of evidence that nonsteroidal anti-inflammatory drugs (NSAIDs) can prevent colorectal cancer. In this article the role of cyclooxygenase 1 and 2, the principle target of NSAIDs, in the development of colorectal cancer is reviewed. Cyclooxygenase is constitutively expressed in normal colonic epithelium and surrounding stroma and could catalyse the generation of malondialdehyde which is a known mutagen and could initiate colorectal carcinogenesis. Mutation of APC which is an early genetic event leads to the expression of cyclooxygenase 2 which may prevents the appropriate apoptosis of mutant adenoma cells. Other proneoplastic effects of cyclooxygenase include changing the action of Transforming Growth Factor beta from anti-proliferative to pro-proliferative, reducing adherence to extracellular matrix, promotes metastasis and angiogenesis. These properties of cyclooxygenases suggest that inhibition of both isoforms may have important protective effects against colorectal cancer.