Antiproliferative and Apoptotic Effects of Zinc-Citrate Compound (CIZAR(R)) on Human Epithelial Ovarian Cancer Cell (OVCAR3) / 대한산부인과학회지

OBJECTIVE: Human seminal plasma has diverse biological activities including cytotoxic effect. It contains high concentrations of zinc and citric acid. Zinc inhibits several carcinoma cell growths through induction of cell cycle arrest and apoptosis. We tried to investigate the effects of zinc-citrate compound (CIZAR(R)) on normal human ovarian epithelial (NOSE) cells and human epithelial ovarian cancer cells, OVCAR-3. METHODS: To investigate the potential effect of CIZAR(R) on cell growth and survival, cells were treated with different dose and exposed to different time. Mitochondrial(m)-aconitase activity was determined in cell extracts using aconitase assay. The flow cytometric assay, DNA laddering, telomerase activity and morphological analysis were done to investigate apoptosis of OVCAR-3 cells. Molecular mechanism of apoptosis was investigated by p53, Bcl-XL, Bcl-2, Bax protein, and caspase activity. RESULTS: Treatment of OVCAR-3 cells with CIZAR(R) resulted in a time- and dose-dependent decrease in cell number in comparison with NOSE cells. M-aconitase activity was significantly decreased in OVCAR-3 cells but relatively constant in NOSE cells. The flow cytometric assay, DNA laddering and morphological analysis indicated apoptosis of OVCAR-3 cells. CIZAR(R) did not affect p53 but increased the expression of p21waf1 upon the indicated times and induced reduction of telomerase activity. CIZAR(R) reduced expression of Bcl-2 and Bcl-xL proteins but induced expression of Bax protein. CIZAR(R) induced apoptosis of OVCAR-3 cells by activation of caspase-3 pathway. CONCLUSION: These results show that CIZAR(R) prevent the proliferation of OVCAR-3 cells by inactivation of m-aconitase activity and induce apoptosis by induction of apoptotic genes and repression of antiapoptotic genes without adverse effect on normal ovarian epithelial cells. These results will offer new window in prevention and treatment of epithelial ovarian cancer.

Collagen-induced Activation of MMPs ( Membrane -type Matrix Metalloproteinase and Matrix Metalloproteinase-2) in ovarian cancer cell lines in Vitro / 대한산부인과학회잡지

OBJECTIVE: Metastatic dissemination of epithelial ovarian carcinoma is thought to be mediated via tumor cell exfoliation into the peritoneal cavity, followed adhesion to and invasion through the mesothelium which overlies the contents of the peritoneal cavity. MMP-2 is secreted as a zymogen, the activation of which has been associated with metastatic progression in human ovarian cancer cell lines. METHODS: We have utilized short-term cultures to analyze the effect of specific extracellular matrix proteins, type I collagen. RESULTS: Culturing Caov-4 ovarian cell line on type I collagen led to a significant increase in conversion of the MMP-2,72kD to the MMP-2,66kD, and MT-MMP expression. MT-MMP expression correlates with expression and activation of MMP-2 during malignant progression. Altered MT-MMP expression in ovarian cell lines might contribute to MMP-2 activation, which facilitates invasion of these tumors. CONCLUSION: In summary, we found increased expression of MT-MMP that correlated with increased level of activated MMP-2 and cellular counts in chemoinvasion assay in Caov-3 cell line. But no significant increases in Skov-4 cell line on type I collagen. Conclusion: These data suggest that type I collagen induces MMP-2 activation in part by up-regulation of MT-MMP expression but has a more complicated mode of action involving additional processes.

Influence of Hypoxic Condition on Invasion of Cultured Trophoblast / 대한주산의학회잡지

OBJECTIVE: To investigated whether lowering oxygen tension affects invasion of cultured trophoblast. METHODS: Trophoblasts were isolated from the normal placenta in early pregnancy(6-10 weeks in gestation). Isolated trophoblasts were cultured under normoxic(5% CO2, 95% humid air in incubator) and hypoxic(MERCK, 1% O2, 99% CO2) conditions for 24, 48 and 72 hours, respectively. The proliferation ability was measured using [H3] thymidine assay. Total RNA was extracted from the cultured trophoblasts. The expressions of matrix metalloproteinase(MMP-2) and tissue inhibitor of metallo- proteinase(TIMP-2) were determined by reverse transcription- polymerase chain reaction(RT-PCR) and Northern blot analysis. The invasiveness of cultured trophoblast was observed using in vitro invasion assay. RESULTS: [H] thymidine assay indicated that cellular DNA synthesis was not affected by the culture condition. The expression of MMP-2 mRNA was decreased at 24 hours and then progressively increased in the time-dependent manner in each culture condition. The expression of TIMP-2 was decreased in the time-dependent manner under hypoxic condition. In vitro invasion assay revealed that the cultured trophoblasts under hypoxic condition has more invasive ability than them under normoxic condition. CONCLUSION: These data suggests that hypoxic condition may stimulates the invasion of trophoblast in the human placentation. And MMP-2 and TIMP-2 may be related to control their invasiveness under hypoxic condition.

Clinical Significance of Zygosity as a Prognostic Factor in Complete Hydatidiform Mole / 대한산부인과학회잡지

To evaluate the possible correlation between the origin of complete hydatidiform mole(CHM) and subsequent persistent gestational trophoblastic tumor(GTT) after molar evacua-tion, we have studied genetic origin patterns against conventional clinical parameters -pati-ent's age, gestational age, uterine size for gestational age, serum beta-hCG levels before mol-ar evacuation- in 69 patients with CHM. In our study, each of large uterine size for gesta-tional age, serum beta-hCG levels before molar evacuation, and genetic origin of CHM had a prognostic significance of subsequent persistent GTT. However, each of gestational age and patient's age is not a good prognostic indicator for subsequent persistent GTT. Among the patients with persistent GTT, there are no differences in clinical parameters- patient's age, gestational age, tumor age(the interval between evacuation of CHM and initiation of chem-otherapy), serum beta-hCG levels before molar evacuation and before initiation of chemother-apy- according to the origin of CHM. There are no differences in the analysis of sex-chr-omosome and variable number tandem repeat sequence YNZ22 and APOB gene in the extr-acted DNA from frozen tissues and paraffin blocks and from EDTA treated peripheral blood and dried blood specimen on Wartman paper. It is suggested that analysis of sex-chromo-some and polymorphism of YNZ22 and APOB gene from the extracted DNA of paraffin bl-ock and dried blood specimen on Wartman paper is the valauble experiment to evaluate the origin and the classification of hydatidiform mole and seems to be the sensitive molecular genetic method in predicting subsequent persistent GTT.