【摘要】 目的研究二甲双胍对人胚肾细胞系HEK293T增殖的抑制和诱导凋亡的作用,并初步探讨其可能的机制。方法采用0mmol/L、5mmol/L、10mmol/L、20mmol/L二甲双胍浓度梯度处理HEK293T细胞,各组分别处理24、48、72h后,通过MTT实验分析细胞增殖变化;在二甲双胍浓度梯度处理48h后,利用Annexin V-FITC/PI染色检测细胞凋亡;采用实时定量PCR检测Cyclin D1基因mRNA的表达变化。结果二甲双胍处理后,所有处理组在24、48、72h的细胞存活率均显著下降(P<0.05),但并未发现有时间依赖和浓度依赖效应;经流式细胞仪检测,二甲双胍干预48h之后,5mmol/L、10mmol/L和20mmol/L浓度处理组HEK293T细胞凋亡率均明显高于对照组(P<0.05);0mmol/L、5mmol/L、10mmol/L、20mmol/L二甲双胍分别处理HEK293T细胞48h后,实时定量PCR结果显示,10mmol/L和20mmol/L处理组的Cyclin D1表达量出现明显降低(P<0.05),但5mmol/L处理组未发现有明显变化(P>0.05)。结论二甲双胍对HEK293T细胞增殖有明显抑制作用,而对HEK293T细胞凋亡有明显的促进作用,这可能是通过抑制细胞内Cyclin D1表达来实现的。更多还原

【Abstract】 Objective To investigate the effects of metformin onhuman embryonic kidney cells HEK293 T and its possible mechanism. Methods The HEK293 T cells were treated with 0 mmol/L,5 mmol/L,10 mmol/L and 20 mmol/L metformin treatment,respectively,for 24,48 and 72 h and performed MTT to analyze the cell proliferation;after 24 h treatment using different concentration of metformin,the cells were stained using Annexin V-FITC/PI to detect cell apoptosis;the cells were treated by different concentration of metformin for 48 h and analyzed by real-time PCR to detect mRNA expression of Cyclin D1. Results After metformin treatment,cell proliferations were significantly decreased in all the treatment groups at 24,48 and 72 h(P<0.05),however,no timedependent and dose-dependent were found;Using the flow cytometry,we found that the 5 mmol/L,10 mmol/L and 20 mmol/L metformingroups had higher apoptosis rates compared to the 0 mmol/L after 48 h treatment;after 48 h treatment of metformin on HEK293 T cells,the real-time PCR results showed that10 mmol/L and 20 mmol/L groups had significant lower Cyclin D1 expression compared to control(P<0.05),however,no significantdifference could be found between 5 mmol/L and control(P>0.05). Conclusion Metformin inhibited HEK293 T cell proliferation and induced HEK293 T cell apoptosis which may achieve via suppressing the Cyclin D1 expression.更多还原