胰岛素样生长因子-1调控下染料木黄酮对睾酮诱导RWPE-1细胞增生的抑制作用

Inhibition of genistein on the proliferation of RWPE-1 cells induced by testosterone under the control of insulin-like growth factor-1

目的研究在胰岛素样生长因子(IGF-1)调控下,染料木黄酮(Gen)对睾酮诱导的人前列腺上皮细胞RWPE-1增生的抑制作用及对细胞周期影响的分子机制。方法采用睾酮刺激RWPE-1细胞增生,观察细胞形态学变化,用0.63、1.25、2.5、5、10、20、40和80μmol/L的Gen与0.1、0.2、0.4、0.8、1.6、3.2、6.4和12.8nmol/L的IGF-1处理RWPE-1细胞24h、48h、72h后,测定各组细胞存活率;流式细胞仪检测对照组、40μmol/LGen组、6.4nmol/LIGF-1组和40μmol/LGen+6.4nmol/LIGF-1组的细胞周期变化,WesternBlot检测细胞周期蛋白cyclinB1、cyclinD1和细胞周期蛋白激酶CDK4。结果 2μmol/L睾酮可显著诱导RWPE-1细胞增生,2.5～80μmol/LGen均能抑制其增生,且抑制作用随着浓度增高和时间延长而增强,6.4nmol/LIGF-1调控下40μmol/LGen可显著减少G0/G1期细胞百分比,增加G2/M期细胞百分比,上调cyclinB1的表达,下调cyclinD1、CDK4的表达。结论 IGF-1调控下Gen可影响细胞周期素和周期素依赖激酶的表达,诱导G2/M期细胞阻滞,抑制睾酮诱导的RWPE-1细胞增生。

Objective To observe the inhibition of genistein （Gen） on the proliferation of RWPE-1 cells under insulin-like growth factor-1（IGF-1） modulation and the molecular mechanism of influences on cell cycles. Methods The proliferation of RWPE-1 cells was promoted by testosterone and assessed by morphologic observation. RWPE-1 cells were treated with Gen of 0. 63 - 80μmol /L and IGF-1 of 0. 1 - 12. 8nmol /L for 24h,48h and 72h, and the viability of the cell was examined. The cell cycle of RWPE-1 in the control group,40μmol /L Gen group, 6. 4nmol /L IGF-1 group and 40μmol /L Gen ＋ 6. 4nmol /L IGF-1 group was observed by flow cytometry analysis. The expression of cyclin B1 ,cyclinD1 and cyclin-dependent kinase 4 was examined by Western blot. Results The proliferation of RWPE-1 cells could be induced by 2μmol /L of testosterone,and the proliferation could be inhibited by 2. 5 - 80μmol /L of Gen. The inhibitive effect could be enhanced with increased concentration and prolonged action time. The percentage of cells at G0 /G1 phase could be decreased and the percentage of cells both at S phase and G2 /M phase could be increased,the expression of cyclinD1 and CDK4 was down-regulated and the expression of cyclinB1 was up-regulated by the treatment of 40μmol /L Gen under the modulation of 6. 4nmol /L IGF-1. Conclusion Under the modulation of IGF-1,Gen could affect the expression of cyclins and CDKs,induce G2 /M cell phase arrested and inhibit the proliferation of RWPE-1 cells induced by testosterone.