RNAi沉默WT1基因对人肝癌细胞HepG2的影响

Effect of RNAi silencing of theWT1 gene on the apoptosis of human hepatocellular carcinoma cell hepG2

目的：探讨利用RNA干扰沉默WT1基因对人肝癌细胞HepG2细胞生长，凋亡的影响，为肝癌的基因治疗提供理论依据。方法：利用脂质体将siRNA真核表达载体转入HepG2细胞中，并检测转染效率；利用免疫细胞化学染色方法检测HepG2细胞中WT1蛋白水平的表达，测定基因沉默效果；用MTT法测定HepG2细胞生长曲线；电镜下观察细胞凋亡形态，流式细胞仪检测凋亡率。结果：利用脂质体为载体将siRNA真核表达载体转染HepG2细胞，转染率达70％以上，转染后细胞中WT1蛋白表达水平下降；RNA干扰沉默WT1基因可抑制细胞增殖，促进细胞凋亡。结论：靶向WT1的序列特异性siRNA可显著抑制WT1基因的表达；下调HepG2细胞WT1基因表达可抑制细胞增殖，促进细胞凋亡。

AIM： To inhibit the expression of WT1 gene in HepG2 cells by siRNA, to study the effect of WT1 on the proliferation and apoptosis. METHODS： HepG2 cells was transfected with WT1-siRNA by liposomes. The protein expression of WT1 in HepG2 cells was detected by immunocytochemistry to detect the effect of WT1 gene silencing. Tumor cells growth was detected by MTT assay. Morphology of apoptotic cells was observed by electron microscope and fluorescence. RESULTS： HepG2 cells was transfected with WT1-siRNA by liposomes, and the expression of WT1 were inhibited signifycantly. The transfection rate was over 70%. Silencing of WT1 by siRNA can inhibit cell proliferation and promote apoptosis. CONCLUSION： Sequence-specific siRNA targeting to WT1 can inhibit WT1 gene expression significantly. The decreesed expression of WT1 gene inhibits cell proliferation and promotes apoptosis of hepG2 cells.