【作者】 刘晓梅；

【导师】 郑桂英；

【作者基本信息】 吉林大学 ， 临床医学， 2010， 硕士

【摘要】 【目的】研究卵泡刺激素（FSH）对卵巢癌细胞株的增殖作用,探讨FSH在卵巢上皮性肿瘤（OET）发生、发展过程中的潜在生物学作用机制。【方法】用含10%胎牛血清的RPMI1640培养液培养HO8910细胞及SKOV3细胞;采用免疫荧光法检测HO8910细胞及SKOV3细胞卵泡刺激素受体（FSHR）的表达;以0 IU/L的FSH作为对照组,不同浓度FSH分别培养HO8910细胞和SKOV3细胞不同时间后,MTT比色法测定各组吸光度值A,并比较各实验组的增殖指数（增殖指数=实验组吸光度（A）/对照组吸光度（A）×100%）;以最佳FSH浓度为实验组,等量培养液替代作为对照组,分别培养最佳时间,用流式细胞仪分别检测实验组及对照组细胞周期分布。【结果】①HO8910细胞FSHR表达阳性,特异性位于细胞核周及细胞质内;SKOV3细胞FSHR表达阴性;②不同浓度FSH作用于HO8910细胞24h、48h、72h时,其A值均较对照组明显升高,其增殖程度与FSH呈浓度依赖效应,当FSH浓度为40 IU/L培养48h时,细胞增殖指数最高（P <0.05）,其后在直至160 IU/L的FSH浓度范围内,该细胞株的增殖并未继续增加,虽然仍有促进作用,但较40 IU/L时却有所减弱。SKOV3细胞各实验组A值均较对照组略有升高,当FSH为40 IU/L培养48h时细胞增殖指数最高（P <0.05）,但未表现出明显的FSH浓度依赖效应。③FSH浓度为40 IU/L时培养HO8910细胞及SKOV3细胞48h,2种细胞G₀/G₁期比例均减少,S期及G₂/M期比例均增加（P <0.05）。而且FSH对HO8910促进细胞分裂作用较SKOV3更明显,差异有统计学意义（P <0.05）。【结论】通过建立FSH作用于HO8910和SKOV3两种细胞模型,证实了FSH与FSHR均与卵巢癌的增殖相关;FSH对FSHR表达阳性的卵巢癌细胞作用呈剂量效应;FSH通过促进细胞周期时相变化而促进卵巢癌细胞增殖,且当FSHR阳性时细胞周期变化更明显。更多还原

【Abstract】 【Objective】Ovarian cancer represents the most lethal gynecologic malignancy, accounting for 3% of all malignancies in women, the mortality rate has always been high in the world.Although ovarian cancer appears to rank second to endometrial cancer in terms of incidence, mortality from ovarian cancer currently exceeds the combined mortality of endometrial and cervical cancer.The peak incidence of invasive epithelial ovarian cancer is at 56 to 60 years of age. Epithelial ovarian cancer (EOC) comprises approximately 90% of ovarian cancers and is thought to arise from the surface epithelium of the ovary. Approximately 70% of women are not diagnosed until the advanced disease is present. If diagnosed at early stages, the 5-year survival rate is 90% compared with only 25% at later stages of the disease. Therefore, it is important to study the causes and pathogenesis of EOC.In the United States, an annual incidence of Ovarian cancer approached 57.3 per 100,000 women around 75 years old. In contrast, in chinese the incidence rate is 75 per 100,000 women. There are some differences bewteen the United States and chinese women. With the study of ovarian cancer etiology,some Scholars found each additional pregnancy after the first also decreases the risk 10%～16%.;The reduction of ovarian cancer risk in women ever taking OC compared with nonusers is about 30%. According to China’s basic national policy, every woman can only have one opportunity to full-term pregnancy, and many American women almost have many full-term pregnancy. Chinese women most use intrauterine contraception, and U.S. women use oral contraceptives most. Therefore, we infer that low follicle stimulating hormone (FSH) environment may have a protective effect on the occurrence of ovarian cancer.The FSH levels of perimenopausal womenwith ovulation most increase in the majority of cycles. The estrogen levels of postmenopausal women will decrease and induce the release of hypothalamic gonadotropin-releasing hormone, the pituitary is stimulated to release FSH. More than 80% of epithelial ovarian cancers are found in perimenopausal and postmenopausal women. we speculated that there are some certain internal relations between the high FSH environment and the occurrence of ovarian cancer. However, the etiology of ovarian cancer is not clear, the independent role of FSH in the development of ovarian cancer is still confusing. a lot of research needs to be done to define the impact of FSH on ovarian cancer in the future.In this research we study FSH and its receptor on the proliferation of ovarian cancer cell line,then discussed the potential biological mechanisms and function of FSH during the occurrence and development of the ovarian epithelial tumors.【Method】We culture HO8910 cells and SKOV3 cells with RPMI1640 culture medium which contains 10% fetal bovine serum.Then,we assay follicle-stimulating hormone receptor (FSHR) expression of HO8910 and SKOV3 cells By immunofluorescence. We take 0 IU / L of FSH as a control group, assay the proliferation of HO8910 and SKOV3 cells cultured in different concentrations of FSH at different time by Methyl thiazolyl tetrazolium (MTT). Then we culture the cells with the best FSH concentration at the best time, detecte the cell cycle distribution by flow cytometry (Fcm). All data are showed with( x±s)in each group. The results were analyzed by use of Analysis of Variance (ANOVA) and processed by spss11.5.【Results】1. The FSHR expression of HO8910 cell is positive, located in the cytoplasm and around the nucleus; SKOV3 cell is negative.2.①We Culture HO8910 cells with different FSH concentration at 24h,48h and 72h, The absorbance was significantly higher than the control group, the degree of their proliferation are in a concentration dependent effect. when FSH concentration is 40 IU / L, the cell proliferations index are the highest at 48h(P <0.05). Then up to 160 IU / L of FSH, the proliferation of the cells did not increase, although some role in promoting , but lower than 40 IU / L.②We Culture SKOV3 cells with different FSH concentration at 24h,48h and 72h, The absorbance was slightly higher than the control group, when FSH concentration is 40 IU / L, the cell proliferations index are the highest at 48h(P <0.05),but the degree of their proliferation are not in a concentration dependent effect.3. HO8910 and SKOV3 cells are cultured with 40 IU / L of FSH at 48h, G0/Gl-phase fraction are both reduced, S phase and G2 / M phase fraction are both increased (P <0.05), and the effect on HO8910 more obvious than SKOV3 (P <0.05).【Conclusions】FSH and FSHR are associated with the proliferation of ovarian cancer through the establishment of FSH acting on the two cell model. FSH have a dose effect on ovarian cancer cells whth positive FSHR expression. .FSH do play an important role in the promotion of cell cycle phase changes, especially on FSHR-positive cells.更多还原