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目的:探讨在雌激素对前列腺平滑肌细胞(PSMC)生物学效应中,雌激素受体α(ERα)和胰岛素样生长因子1(IGF1)对细胞增殖的影响。方法:构建pGSadeno-ERα腺病毒载体,体外转染原代培养的小鼠PSMC后,以半定量逆转录PCR和Western印迹分别检测ERαmRNA和蛋白质的表达;成功获取ERα下调细胞后,加入10-8mol/L17β雌二醇,6h后Western印迹检测细胞IGF1的表达;以噻唑蓝比色分析法(MTT法)评价转染后雌激素或外源性IGF1对PSMC的增殖活性。结果:加入17β雌二醇后,正常对照组PSMC增殖明显,IGF1基因表达水平明显增高(P<0.05);而在ERα下调细胞组中,PSMC增殖、IGF1基因表达水平均无明显变化(P>0.05)。外源性IGF1作用于ERα下调的PSMC后,未能激活细胞增殖(P>0.05)。结论:雌激素在对PSMC的生物学效应中,通过ERα的介导刺激产生IGF1,并且IGF1在ERα的共同参与下促进细胞的增殖。
Objective: To investigate the effects of estrogen on the proliferation of prostatic smooth muscle cells (PSMCs) in vitro and in vitro. Methods: The pGSadeno-ERα adenovirus vector was constructed and transfected into primary cultured mouse PSMC in vitro. The expression of ERαmRNA and protein was detected by semi-quantitative reverse transcription PCR and Western blot respectively. After ERα down-regulated cells, 10-8mol / L17β estradiol. The expression of IGF1 was detected by Western blotting 6h later. The proliferative activity of transfection estrogen or exogenous IGF1 on PSMC was evaluated by MTT assay. Results: After adding 17β-estradiol, PSMC proliferation in normal control group was significantly increased and IGF1 gene expression was significantly increased (P <0.05), while in ERα-regulated cells, PSMC proliferation and IGF1 gene expression did not change significantly (P> 0.05). Exogenous IGF1 could not activate cell proliferation after ERα downregulated PSMC (P> 0.05). CONCLUSIONS: Estrogen produces IGF1 through ERa-mediated stimulation of the biological effects of PSMC, and IGF1 promotes cell proliferation with the involvement of ERα.