下调PAR基因表达对前列腺癌PC3细胞周期及Cdc25C蛋白表达的影响

摘要

背景与目的:前列腺雄激素调节基因(prostate androgen regulated gene,PAR)在雄激素非依赖性前列腺癌中特异性高表达,可能与细胞恶性转化相关.本研究用RNA干扰技术下调PAR基因表达,并探讨其对前列腺癌PC3细胞细胞周期及调节因子Cdc25C表达的影响.方法:针对PAR基因的siRNA转染PC3细胞,以RT-PCR验证其对PAR基因表达抑制的有效性,用流式细胞术检测细胞周期,Western blot检测Cdc25C表达水平.结果:siRNA转染可抑制PC3细胞PAR基因表达,同时G2/M期细胞及凋亡细胞比例分别由(23.79±3.16)％及(1.49±0.13)％增加至(29.95±3.25)％和(20.61±2.73)％,Cdc25C蛋白表达水平明显下降.结论:PAR基因表达下调可诱导细胞G2/M期阻滞和凋亡,抑制细胞周期调节因子Cdc25C的表达.%Background and purpose: The expression of PAR gene was higher in androgen-resistant prostate cancer (Pca) in comparison to androgen-sensitive Pca and may be involved in the cellular malignant transformation. This study was designed to investigate the effects of downregulated PAR expression on the cell cycle of PC3 cells and to explore the role of Cdc25C in this process. Methods: Suppression of PAR expression was achieved by transfection of PC3 cells with small interfering RNA (siRNA) target against PAR. Phase distribution of cell cycle was analyzed by flow cytometry. Western blot was performed to investigate the expression of Cdc25C. Results: The expression of PAR was suppressed by siRNA. Flow cytometry revealed a cell cycle G2/M arrest and induction of apoptosis. Western blot showed the expression of Cdc25C decreased after PAR expression downregulated. Conclusion: Downregulated PAR expression inhibits the expression of Cdc25C and thus is involved in the G2/M arrest and apoptosis of PC3 cells.