The Expressions Of GnRHⅠ And GnRHR In Epithelial Ovarian Tumor And GnRH Agonist Effect On Ovarian Cancer

Ovarian cancer is the highest mortality in gynecological malignancies, but the mechanism by which ovarian cancer occuring is unclear. The results of a large number of epidemiological investigations have proved that high gonadotropin levels and excessive stimulation of the ovulation could induce ovarian cancer. Gonadotropin-releasing hormone (GnRH) can bind to its cell membrane receptor specifically and regulate the formation of receptors in gonads and the functions of hormone. There are two different isoforms of GnRH receptor in the mammals'gonads, including GnRHⅠandⅡreceptor, which have different function . Substantial evidence showed that GnRH and its receptor can be widely found in hormone- dependent neoplasms, for example, breast carcinoma,ovarian cancer,endometrial carcinoma,carcinoma of prostate,liver cancer, GnRH and GnRH receptor can control the growth of tumor. GnRH analogues is a kind of high-performance and harmless casualty agent for hormonal dependent tumor. Therefore, further exploring of the role and mechanisms of GnRH and its receptor will provide valuable evidence for targeting therapy on ovarian cancer. In order to clarify the role and mechanisms of GnRH and its receptor, the expression of GnRH and its receptor in benign and malignant tumors were first detected. Meanwhile, To investigate GnRH agonist inhibit the growth of ovarian carcinoma cells, we can get the mechanism of increasing chemotherapy and provide experimental evidences about treating ovarian cancer.PartⅠExpressions of GnRHⅠand GnRH receptor in Ovarian tumorObjective:To study the expressions of GnRHⅠand GnRH receptor in Epithelial Ovarian Cancer and their differences, and get the results about the relation between expressions of GnRHⅠand GnRH receptor and pathological differentiation.Methods:The expressions of GnRHⅠand GnRH receptor in ovarian benign tumor(22 cases) and epithelial ovarian carcinoma(39 cases were detected by immunohistochemical.Results:1 The localization and distribution of GnRHⅠand GnRH receptor: The localization of GnRHⅠpositive product is in the cytoplasm or cell membrane, no positive signals around the cell nucleus. The localization of GnRH receptor positive product is in the cytoplasm or cell membrane, no positive signals around the cell nucleus.2 Epithelial ovarian tumor in the expression of GnRHⅠand its relationship with differentiation: The expression rates of GnRHⅠreceptor in benign and malignant tumors were 54.5% and 79.5%, the expressions of GnRHⅠreceptor in ovarian benign tumors was significantly lower than that in epithelial ovarian cancer (X2=4.21,P<0.05), the results have significant differences. The positive expression levels of GnRHⅠreceptor in well-differentiated ovarian carcinoma (50%) have significantly lower than middle-differentiated ovarian carcinoma (91.7%), (X2=4.44, P<0.05); the positive expression levels of GnRH receptor in poor-differentiated ovarian carcinoma (93.3%) have no significant differences than middle differentiated ovarian carcinoma (91.7%) (X2=0.03, P>0.05).3 Epithelial ovarian tumor in the expression of GnRH receptor and its relationship with differentiation:The expression rates of GnRH receptor in benign and malignant tumors were 81.8% and 53.8% ,the expressions of GnRH receptor in ovarian benign tumors was significantly higher than that in epithelial ovarian cancer (X2=4.77,P<0.05), the results have significant differences. The positive expression levels of GnRH receptor in well-differentiated ovarian carcinoma (16.7%) have significantly lower than middle-differentiated ovarian carcinoma (58.3%),(X2=4.44,P<0.05); the positive expression levels of GnRH receptor in poor-differentiated ovarian carcinoma (80.0%) have no significant differences than middle-differentiated ovarian carcinoma (58.3%), (X2=1.50, P>0.05).Conclusion:1 The changes of GnRH I and GnRH receptor protein expression in ovarian epithelial tumors, prompt the role of GnRH autocrine/paracrine regulatory system were involved in the development of ovarian epithelial malignant tumors.2 The expression of GnRHⅠand GnRH receptor was related to pathological differentiation level. As the pathology of the higher level, the rate of the expression GnRH I and GnRH increased, we suggested that GnRH I and GnRH receptor relate to the growth of ovarian cancer, invasiveness and prognosis.Part 2 The effects of GnRH agonist triptorelin on human ovarian cancer cellsObjective:Observe the inhibition about Triptorelin efforts to OVCAR3 cell and discuss the mechanism of apoptosis.Methods:1 Prolifferation activities of OVCAR3 cells treated with or without Triptorelin were detected by MTT assay2 Expression level of Caspase-3 in OVCAR3 cells treated with or without Triptorelin were assayed by RT-PCR.Results:1 Triptorelin effected the growth of OVCAR3: By the met-hod of MTT, we found that when triptorelin in 10-7mol/l, 10-6mol/l, 10-5mol/l, 10-4mol/l, 10-3mol/l, 10-2mol/l con -centration effected on OVCAR3 for 24h, the rate of inhibition is (41.07±0.43)%,(50.56±0.76)%,(69.86±1.05)% (65.43±0.87)%,(62.09±1.24)%,(58.69±0.87)%, when triptor-elin in 10-7mol/l concentration, cells growth are significa-ntly inhibited, have statistical significance (P<0.05); wh-en triptorelin in 10-5mol/l concentration effect cells 24h, 48h, 72h, the rate of inhibition is (69.86±1.08)%,(58.69±0.98)%,(48.56±1.02)%, when triptorelin effected on OV-CAR3 for 24h, cells growth are significantly inhibited, have statistical significance(P<0.05).2 Study of Caspase-3 related apoptosis when triptorelin effect on cells by using the method of RT-PCR:①when triptorelin in 10-4mol/l,10-5mol/l,10-6mol/l concentration effected on OVCAR3 for 24h, the OD of Caspase-3 is (0.51±0.04), (0.71±0.07),(0.41±0.04), compared with the contrl group that the OD of Caspase-3 is (0.48±0.04), the highest expression of Caspase-3 when triptorelin in 10-5mol/L concentration effected on OVCAR3, have statistical significance (F=42.028,P<0.05);②when triptorelin in 10-4mol/l, 10-5mol/l,10-6mol/l concentration effected on OVCAR3 for 48h, the OD of Caspase-3 is (0.69±0.07), (1.01±0.08), (0.64±0.08), compared with the contrl group that the OD of Caspase-3 is (0.48±0.04), the highest expression of Caspase-3 when triptorelin in 10-5mol/L concentration effected on OVCAR3, have statistical significance (F=107.595,P<0.05);③when triptorelin in 10-4mol/l concentration effected on OVCAR3 for 24h and 48h, the OD of Caspase-3 is (0.85±0.09), (0.62±0.06), compared with the contrl group that the OD of Caspase-3 is (0.34±0.04), the highest expression of Caspase-3 when triptorelin effected on OVCAR3 for 24h, have statistical significance for 24h, have statistical significance(F=47.164,P<0.05);④when triptorelin in 10-5mol/l concentration effected on OVCAR3 for 24h and 48h, the OD of Caspase-3 is (0.85±0.09),(0.64±0.05) compared with the contrl group that the OD of Caspase-3 is (0.40±0.06), the highest expression of Caspase-3 when triptorelin effected on OVCAR3 for 24h, have statistical significance (F=29.647,P<0.05);⑤when triptorelin in 10-6mol/l concentration effected on OVCAR3 for 24h and 48h, the OD of Caspase-3 is (0.96±0.08),(0.68±0.04) compared with the contrl group that the OD of Caspase-3 is (0.58±0.04), the highest expression of Caspase-3 when triptorelin effected on OVCAR3 for 24h, have statistical significance (F=48.337,P<0.05).Conclusion:1 Triptorelin can inhibit ovarian carcinoma cells which express GnRH receptor.2 Triptorelin induce antiproliferation and related to Caspase-3 in epithelial ovarian carcinoma cells, after triptorelin effect on OVCAR3, the expression of Caspase-3 increased.