Genistein对人卵巢癌细胞系3AO抑制增殖的作用及机制的探讨

Inhibitory effects of genistein on proliferation in human ovarian carcinoma cell line 3AO and related mechanism

目的:研究植物雌激素染料木黄酮对人卵巢癌细胞系3AO细胞增殖的抑制作用和凋亡的诱导作用,探讨其抗癌作用的机制.方法:应用MTT法检测不同浓度染料木黄酮对3AO细胞的生长抑制作用,电镜观察药物作用后细胞的超微结构改变,TUNEL法确定凋亡,流式细胞仪检测细胞周期、定量分析凋亡,免疫细胞化学技术检测雌激素β受体的表达.结果:3AO细胞经不同浓度染料木黄酮处理后,细胞生长明显受到抑制,且这种抑制作用随时间延长及浓度增高而增强,10mg/L和20mg/L染料木黄酮作用72h后,抑制率分别达到54.24%和65.99%.染料木黄酮阻断3AO细胞生长于G2/M期,在G0/G1前出现典型亚二倍体凋亡峰,且凋亡呈时间依赖性.电镜观察到用药后凋亡细胞典型的形态学特征.TUNEL法观察到大量阳性凋亡细胞.免疫细胞化学法检测到用药后,ERβ表达明显加强.结论:染料木黄酮对3AO细胞的生长有明显的呈时间及浓度依赖性的抑制作用,并诱导其发生凋亡.染料木黄酮可能通过雌激素β受体途径发挥药理作用,诱导卵巢癌细胞发生凋亡.

AIM： To investigate the effects of genistein on proliferation inhibition and apoptosis induction in human ovarian carcinoma cell line 3AO and to explore its anti-cancer mechanism. METHODS： After cultured with different doses of genistein for different time periods, the cells were tested by MTr assay. Study on the ultrastructure of 3AO cells was performed by transmission electron microscope （TEM） after the administration of genistein. Apoptosis was detected by means of terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling （TUNEL） method. Apoptosis rate and cell cycle distribution of 3AO cells were measured by flow cytometry （FCM）. The expression of estrogen receptor β protein was measured by immunocytochemical technique. RESULTS： After 3AO cells were treated with different concentrations of genisteln, dose- and time-dependent growth inhibition was demonstrated. Inhibition percentage in cells exposed to 10 mg/L and 20 mg/L genistein for 72 h were, respectively, 54.24% and 65.99%. Also, genistein could block 3AO cells in the G2/M phase of cell cycle, and a typical sub-diploid apoptosis peak was demonstrated before G1/G2 phase. Moreover the apoptosis was time-dependent. The characteristic morphological changes of apoptosis in genistein-treated 3AO cells were observed by TEM. TUNEL staining revealed apoptosis-positive ceils. Genistein could up-regulate the expression of ERβ. CONCLUSION： Genistein can dose- and time-dependently inhibit growth and induce apoptosis in ovarian carcinoma cell line 3AO the potential pathway of ERβ.