CAJ | 학술논문

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。
목적：연구란포자격소( FSH)대란소암세포주적증식작용,탐토FSH재란소상피성종류( OET)발생、발전과정중적잠재생물학작용궤제。방법용함10%태우혈청적RPMI1640배양액배양HO8910세포급SKOV3세포；채용면역형광법검측HO8910세포급 SKOV3세포란포자격소수체( FSHR)적표체；이0 IU/L적FSH작위대조조,불동농도FSH분별배양HO8910세포화SKOV3세포불동시간후,MTT비색법측정각조흡광도치A,병비교각실험조적증식지수(증식지수=실험조흡광도( A)/대조조흡광도( A)×100%)；이최가FSH농도위실험조,등량배양액체대작위대조조,분별배양최가시간,용류식세포의분별검측실험조급대조조세포주기분포。결과①HO8910세포FSHR표체양성,특이성위우세포핵주급세포질내；SKOV3세포FSHR표체음성；②불동농도FSH작용우HO8910세포24 h、48 h、72 h시,기A치균교대조조명현승고,기증식정도여FSH정농도의뢰효응,40 IU/L FSH배양48 h시,세포증식지수최고(P<0．05),수착FSH농도승고(직지160 IU/L),해세포주적증식병미계속증가。 SKOV3세포각실험조A치균교대조조략유승고,40 IU/L FSH배양48 h시,세포증식지수최고(P<0．05),단미표현출명현적FSH농도의뢰효응；③FSH농도위40 IU/L시,배양HO8910세포급SKOV3세포48 h,량충세포 G0/G1기비례균감소, S 기급 G2/M 기비례균증가( P <0．05)。이차 FSH 대HO8910촉진세포분렬작용교 SKOV3경명현,차이유통계학의의( P <0．05)。결론통과건립 FSH 작용우HO8910화SKOV3량충세포모형,증실FSH여FSHR균여란소암적증식상관；FSH대FSHR표체양성적란소암세포작용정제량효응；FSH통과촉진세포주기시상변화촉진란소암세포증식,차당FSHR양성시,세포주기변화경명현。
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 RPMI1640 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 pro-liferation of HO8910 and SKOV3 cells cultured in different concentrations of FSH was assayed at different time points by Methyl thiazolyltetrazolium (MTT). 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 in-crease 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 pro-liferation 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 SK-OV3 ( 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,espe-cially on FSHR-positive cells.