| Objective:The mechanisms involved in ovarian cancer tumourigenesis are not fully understood,one hypothesis that reproductive hormone may be involved in carcinogenesis and development of ovarian cancer are still controversial. The study is to observe the effect and the possible mechanism of reproductive hormones treated on cell lines,to provide the theoretical foundation for explore clinical effection of reproductive hormones for the carcinogenesis development and prognosis of Ovarian cancer, and the clinical endocrine therapy for Ovarian cancer.Methods:Ovarian cancer cell line SKOV3,OVCAR-3are treated by different does of estrogen,progesterone and human chorionic gonadotropin, then use various methods to detect the growth condition:(1) Estimating the cell proliferation by3-(4,5-dimethylthiazol-z-yl)-2,5-dipheny tetrazolium blue (MTT) colorimetric assay after24,48,72houes treatment.(2) Analyzing the the changing of distribution of cell cycle and early apoptosis by flow cytometry after hormones treatment;(3) The cell lines estrogen receptors (ERa, ERβ) mRNA, progesterone receptors(PRA, PRB) mRNA,human chorionic gonadotropin receptor (LHCGR) mRNA expressions and after72h treatment of different kinds of hormones the expressions of EGFR,Bcl-2,Bax mRNA in SKOV3cell are detected by real-time PCR.Results:(1) P and E of10-4mol/L inhihibit the proliferation of SKOV3、OVCAR-3cell lines, and the inhibition depends on the exposure time and dose;after incubate with HCG and E of concentration≤10-6mol/L, cell proliferation increased significantly, the proliferation depends on the exposure time.(2) The analysis of cell cycle showed that:72h incubation with10-4mol/L P blocked cells at G0/G1phase,while72h incubation with10-4mol/L E blocked cells at S phase, then after72h treatment with500U/L HCG and10-6mol/L E cell percentage in G2/M phase of cell cycle increased slightly.(3) The analysis of cell apoptosis showed that:P and E of10-4mol/L can induce significantly apoptosis compare with control group (P<0.05), the apoptosis ratio are20%and50%;(4) Cell line SKOV3has a high level of Era, is more sensitive to E compared with cell line OVCAR-3which Erα level is low; the ihibitit effect of P is more significent to cell line OVCAR-3,which has a higher PRA:PRB ratio than SKOV3; The proliferation of cell line OVCAR-3, which has a high level of LHCGR, and cell line SKOV3which has lower level, are all increased significantly compared with control group, while OVCAR-3cell increased more significantly(P<0.01); But after72h incubation, both of the cell lines’proliferation increased significantly (P<0.01). After72h treatment with10-4mol/L P and E the expressions of EGFR, Bcl-2mRNA are decreased significantly (P<0.01),after incubate with HCG and10-6mol/L E the expressions of EGFR,Bcl-2mRNA increased significantly (P<0.05).Conclusion:P and high concentration of E could inhibit the proliferation of ovarian cancer cell, the effects were achieved by apoptosis related to the blocking of cell cycle at G0/G1and S phase, down-regulation of the expressions of EGFR and Bcl-2; HCG and lower concentration of E could stimulate the proliferation of ovarian cancer cells, the effects were achieved by up-regulation of the expressions of EGFR and Bcl-2. The effects of E to ovarian cancer cells with could be mainly mediated through interaction with Era, while P could be mainly mediated through interaction with PRB, and then interaction with LHCGR is not the only way which could effect ovarian cancer cells. |
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