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目的观察沉默信息调节因子1(SIRT1)小干扰RNA(si RNA)对前列腺癌细胞PC3细胞生长增殖、DNA合成、细胞凋亡和Bcl-2和Bax蛋白的表达变化,探讨SIRT1在前列腺癌发生中的可能机制。方法体外培养PC3细胞,分空白对照组(mock组),转染阴性对照组(scramble si RNA组)和SIRT1 si RNA转染组;Western blot检测PC3细胞中SIRT1的干涉效能;MTT法测定PC3细胞的增殖率;Brd U掺入法测定DNA合成;流式细胞术检测细胞凋亡;Western blot检测PC3细胞中细胞凋亡关键调控因子Bcl-2和Bax的蛋白表达。结果与对照组比较,SIRT1 si RNA组SIRT1蛋白表达降低(P<0.01),PC3细胞的增殖和DNA合成明显受抑制(P<0.01),细胞凋亡比例增加(P<0.01),Bcl-2蛋白表达减少,Bax的表达增加。结论下调SIRT1的表达抑制细胞增殖和DNA合成,诱导前列腺癌PC3细胞发生凋亡,其机制可能与改变细胞凋亡关键调控因子Bcl-2和Bax的蛋白表达相关。
Objective To observe the changes of proliferation, DNA synthesis, apoptosis and the expressions of Bcl-2 and Bax proteins in PC3 cells of prostate cancer cells by SIRT1 siRNA (small interfering RNA (siRNA)), and to explore the role of SIRT1 in the pathogenesis of prostate cancer Possible mechanism. Methods PC3 cells were cultured in vitro, and divided into blank control group (mock group), transfected negative control group (scramble si RNA group) and SIRT1 si RNA transfected group. The interference effect of SIRT1 in PC3 cells was detected by Western blot. . The DNA synthesis was measured by BrdU incorporation method. The apoptosis of PC3 cells was detected by flow cytometry. The protein expressions of Bcl-2 and Bax, the key regulatory factors in PC3 cells, were detected by Western blot. Results Compared with the control group, SIRT1 protein expression in SIRT1 si RNA group was significantly decreased (P <0.01), PC3 cell proliferation and DNA synthesis were significantly inhibited (P <0.01) Protein expression decreased, Bax expression increased. Conclusion Down-regulation of SIRT1 expression can inhibit cell proliferation and DNA synthesis and induce apoptosis of PC3 cells. The possible mechanism may be related to the alteration of the protein expression of Bcl-2 and Bax, the key regulatory factors of apoptosis.