c-Jun基因RNA干扰对大鼠血管平滑肌细胞增殖的影响

Effect of RNA Interference for c-Jun Gene on Proliferation of Rat Vascular Smooth Muscle Cells

目的以RNA干扰抑制血管平滑肌细胞(vascular smooth muscle cells,VSMCs)的c-Jun基因表达,探讨其表达抑制后VSMCs增殖的变化。方法设对照组(VSMCs不作处理)、阴性si RNA组(VSMCs转染无关序列的si RNA)及c-Jun si RNA组(VSMCs转染c-Jun si RNA)。应用RT-PCR半定量法检测VSMCs中c-Jun的mRNA水平,Western blot法检测VSMCs中c-Jun的蛋白水平,应用MTT比色法及3H-TdR掺入法检测VSMCs的增殖情况,流式细胞仪检测VSMCs的细胞周期变化。结果c-Jun si RNA组的c-Jun mRNA及蛋白表达水平均较对照组明显降低(P<0.05,P<0.01),而阴性si RNA组与对照组间差异无统计学意义(P>0.05)。c-Jun si RNA组VSMCs的增殖活性较对照组明显降低(P<0.05),细胞周期出现明显的G0/G1期阻滞;而阴性si RNA组与对照组间差异无统计学意义(P>0.05)。结论RNA干扰介导的c-Jun基因沉默可显著抑制VSMCs的体外增殖。

Objective To investigate the influence of RNA interference targeting c-Jun gene on the proliferation of rat vascular smooth muscle cells （VSMCs）. Methods The experiment was performed with c-Jun siRNA （c-Jun siRNA group）, control reverse sequence siRNA （control siRNA group） or no siRNA （control group）. VSMCs were transfected with siRNA targeting c-Jun gene by liposome. Effects of c-Jun siRNA on mRNA and pro rein expressions of c Jun were examined by RT-PCR analysis and Western blot respectively. MTT test and 3 H-TdR incorporation were used to detect VSMCs proliferation. Cell cycle analysis of VSMCs in vitro was determined by flow cytometer. Results The expression levels of mRNA and protein of c Jun in c-Jun siRNA group were significantly lower than those in control group （P〈0.05, P〈0.01）. There was no significant difference between control group and control siRNA group （P〉0.05）. Proliferation activity of VSMCs decreased significantly in c Jun siRNA group compared with that in control group （P〈0.05） and VSMCs was blocked in the G0/G1 phase of cell cycle significantly （P〈0.05）. There was no significant difference between control group and control siRNA group （P〉 0.05）. Conclusion c-Jun gene silenced by RNA interference can inhibit VSMCs proliferation effectively in vitro.