丙戊酸钠对人肾癌ACHN细胞增殖的抑制作用及其机制探讨

Inhibitory effect of valproate on proliferation of human kidney carcinoma ACHN cells and its mechanism

目的:探讨丙戊酸钠(valproate,VPA)对人肾癌ACHN细胞的增殖、细胞周期及凋亡的影响及其可能机制。方法:采用CCK-8法观察VPA对肾癌ACHN细胞的增殖抑制作用。FCM法检测VPA对肾癌ACHN细胞周期和细胞凋亡的影响。实时荧光定量PCR法检测细胞周期调节因子cyclin E1和P^21WAF1以及凋亡调控因子Bcl-2和Bax的m RNA表达。结果:不同浓度的VPA作用于肾癌ACHN细胞48h后均能显著抑制细胞增殖,其半数抑制浓度(50% inhibitory concentration,IC_50)为4.21mmol/L。VPA处理组与对照组相比,G_0/G_1细胞所占比例增加,细胞凋亡率明显增加。4mmol/LVPA处理细胞48h后,cyclin E1 mRNA表达水平明显降低,P^21WAF1mRNA表达量明显增加,Bcl-2mRNA表达水平明显降低,Bax mRNA表达量明显增加。结论:VPA通过诱导细胞周期阻滞和细胞凋亡,抑制肾癌ACHN细胞增殖。

Objective： To investigate the effects of VPA （valproate） on proliferation, cell cycle distribution and apoptosis of human kidney carcinoma ACHN cells and the possible underlying mechanisms. Methods：The effect of VPA on the proliferation of ACHN cells was examined by CCK-8 （cell counting kit-8） assay. Flow cytometry was used to analyze the cell cycle distribution and apoptosis of ACHN cells exposed to VPA. The mRNA expressions of cyclin E1, P^21WAF1, Bcl-2 and Bax were detected by real-time fluorescence quantitative-PCR. Results： Incubation with various concentrations of VPA for 48 h resulted in a significant inhibition of proliferation of ACHN cells with an IC50 （50% inhibitory concentration） value of 4.21 mmol/L. After treatment with VPA, the cell cycle was arrested obviously at G0/G1 phase and the apoptotic rate was significantly increased as compared with the control group. After treatment with 4 mmol/L VPA for 48 h, the levels of P^21WAF1 and Bax mRNAs were both significantly increased, and at the same time, the levels of cyclin E1 and Bcl-2 mRNAs were obviously decreased. Conclusion： VPA can inhibit the proliferation of kidney carcinoma ACHN cells by inducing cell-cycle arrest and apoptosis.