siRNA-ID-1对人肝癌细胞HepG2中ERK信号通路的影响

The Effects of siRNA-ID-1 in the ERK1/2 Signaling Transduction Pathway of HepG2 Cell

目的观察si RNA-ID-1转染人肝癌细胞HepG2细胞后对肝癌细胞系HepG2细胞增殖的影响及其对ERK1/2通路的作用。方法实验分为4组,即空白对照组、转染试剂组、转染对照组及转染si RNA-ID-1组。阳离子脂质体法介导si-ID-1转染肝癌细胞后,唑蓝比色法(MTT)观察HepG2细胞增殖情况。分别用反转录聚合酶链式反应法(RT-PCR)和蛋白免疫印迹法(Western blotting)检测转染后p-ERK1/2、ERK1/2 mRNA与蛋白表达的情况。结果转染si RNA-ID-1的HepG2细胞增殖速度明显减慢(P<0.01)。转染后ERK1/2及p-ERK1/2 mRNA表达降低(P<0.01);p-ERK1/2蛋白的表达较空白对照组明显降低(P<0.05),ERK1/2蛋白表达空白变化不明显。结论si RNA-ID-1转染HepG2细胞后可明显抑制细胞的增殖,使得ERK1/2基因表达活性下降,提示ID-1有可能通过ERK1/2信号转导通路介导,促进肝细胞癌HepG2细胞的增殖。

Objective To investigate the effect of and the effects of ERK1/2 signaling transduction pathway siRNA-ID-1 on the proliferation of HepG2 cells on the inhibition of HepG2 cells by siRNA. Methods The experiment divides into four groups, normol control group, Lipofectamine 2000 group,control-siRNA group, Id-1 siRNA group, si-ID-1 was synthesized and transfected into HepG2 cell by Lipo- fectamine 2000. Cell growth was measured with MTT assay. ERK1/2 and p-ERK1/2 mRNA expres- sion were determined by RT-PCR while their protein expressions were measured by using Western blot method. Results The cell proliferation was inhibited when transfected with siRNA-ID-1（P〈0.01）. Compared to that of the control groups, the mRNA level of ERK1/2 and p-ERK1/2 was reduced （P〈0.01）, whereas the protein level of ERK1/2 demonstrated no significant difference（P〉0.05） and p-ERK1/2 was reduced（P〈0.05）. Conclusion RNA interference could inhibit the growth of HepG2 and the expression of ERK1/2 gene, which shows that ID-1 may increase the proliferation rate via ERK1/2 signaling pathway.