卵泡刺激素及其受体对卵巢癌细胞株的增殖作用

Effects of FSH and FSHR on proliferation of ovarian cancer cell line

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,M TT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24 h、48 h、72 h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,40 IU/L FSH培养48 h时,细胞增殖指数最高(P<0.05),随着FSH浓度升高(直至160 IU/L),该细胞株的增殖并未继续增加。SKOV3细胞各实验组A值均较对照组略有升高,40 IU/L FSH培养48 h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48 h,两种细胞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促进卵巢癌细胞增殖,且当FSHR阳性时,细胞周期变化更明显。

Objective To study the effects of FSH and its receptor on the proliferation of ovarian cancer cell line,then discuss the potential biological mechanisms and function of FSH during the occurrence and development of the ovarian epithelial tumors. Methods We cultured HO8910 cells and SKOV3 cells with RPM I1640 culture medium which contained 10% fetal bovine serum. Then the follicle-stimu / lating hormone receptor（ FSHR） expression of HO8910 and SKOV3 cells was assayed by immunofluorescence. 0 IU / L of FSH was taken as a control group,the proliferation of HO8910 and SKOV3 cells cultured in different concentrations of FSH was assayed at different time points by M ethyl thiazolyltetrazolium（ M TT）. Then the cells were cultured with the best FSH concentration at the best time,the cell cycle distribution was detected by flow cytometry（ Fcm）. Results 1The FSHR expression of HO8910 cell was positive,it located in the cytoplasm and around the nucleus; SKOV3 cell was negative. 2At 24 h,48 h,72 h after HO8910 cells with different concentrations of FSH,the absorbance was significantly higher than that of control group,the degree of proliferation was in a concentration dependent manner. When FSH concentration was 40 IU / L,the cell proliferation index was the highest at 48 h（ P〈0. 05）. The proliferation of the cells did not increase,along with the increase of FSH concentration（ up to 160 IU / L） 2At 24 h,48 h,72 h after SKOV3 cells with different concentrations of FSH,the absorbance was slightly higher than that of control group,when FSH concentration was 40 IU / L,the cell proliferation index was the highest at 48 h（ P〈0. 05）,but the degree of proliferation was not in a concentration dependent manner. 3HO8910 and SKOV3 cells were cultured with 40 IU / L of FSH for 48 h,G0/ G1-phase fraction both reduced,S phase and G2/ M phase fraction both increased（ P〈0. 05）,and the effects on HO8910 were more obvious than SKOV3（ P〈0. 05）. Conclusion FSH and FSHR are confirmed to associate with the proliferation of ovarian cancer through the establishment of FSH acting on the HO8910 and SKOV3 cells. FSH has a dose effect on ovarian cancer cells with positive FSHR expression. FSH plays an important role in the promotion of cell cycle phase changes,especially on FSHR-positive cells.