重组人苗勒抑制物对2株卵巢癌细胞增殖的抑制作用

Inhibitory effects of recombinant human Mullerian inhibiting substance on cell proliferation in two human ovarian carcinoma cell lines

目的:探讨重组人苗勒抑制物(rhMIS)对人卵巢癌细胞株OVCAR8及SKOV3细胞生长的影响。方法:Western blotting检测细胞中MISII型受体(MISIIR)蛋白表达,激光共聚焦显微镜观察MISIIR蛋白在细胞中定位。rhMIS分别干预2株卵巢癌细胞,四甲基偶氮唑蓝(MTT)检测细胞增殖变化;软琼脂克隆实验检测细胞体外成瘤性;流式细胞术分析rhMIS对细胞凋亡和细胞周期影响。结果:OVCAR8细胞表达MISIIR蛋白,其定位于细胞表面及细胞质中,SKOV3细胞中MISIIR蛋白表达缺损。经rhMIS作用48h后,OVCAR8细胞的生长速率显著减慢、细胞体外成瘤性明显降低、细胞发生凋亡和G1期细胞增加,rhMIS(10mg/L)干预后细胞凋亡率和G1期细胞比率分别可高达(31.3±2.1)%和(70.4±3.0)%,与SKOV3细胞比较差异显著(P<0.01)。结论:重组人苗勒抑制物可以抑制MISIIR蛋白表达的卵巢癌细胞增殖、诱导其凋亡、阻滞细胞周期,这有望成为治疗卵巢癌的一个新靶点。

AIM： To investigate the inhibitory effects of recombinant human Mullerian inhibiting substance on cell proliferation in human ovarian carcinoma cells （OVCAR8 and SKOV3 cell lines）. METHODS： The expression of MI- SⅡR protein and the localization of MISⅡR protein were analyzed by Western blotting and confocal spectral microscopy, respectively. Cell apoptosis and cell cycle were detected by flow cytometry （FCM）. Cell viability was determined via MTr method. Clone formation test was used to detect oncogenicity in vitro. RESULTS： The MISⅡR protein expression in OVCAR8 cells but not in SKOV3 cells was observed. MISⅡR expression was seen on the OVCAR8 cell surface and in the cytoplasm with both antibodies. After treated with rhMIS for 48 h, the cell viability was significantly decreased in OVCAR8 cells, rhMIS inhibited the oncogenicity of OVCAR8 cells greatly. The cell apoptosis of OVCAR8 cell exposed to 10 mg/L rhMIS was （31.3 ±2. 1 ）% , and OVCAR8 cells in the G1 phase were increased by （70. 4±3.0）%. Compared to SKOV3 cells the differences were significant （P 〈 0. 01 ）. CONCLUSION： Recombinant human Mullerian inhibiting substance suppresses the growth of MISⅡR - positive ovarian cancer ceils by inducing apoptosis and cell cycle arrest. We predict that rhMIS might be a new target to treat human ovarian malignancies.