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目的探讨膜联蛋白A7低表达对人肝癌HepG2细胞的增殖产生的影响。方法采用Western blotting法鉴定siRNA可有效抑制膜联蛋白A7表达,然后用脂质体转染法将siRNA转染入HepG2细胞,将细胞分为siRNA干扰组、阴性对照组和空白对照组,在转染后24h、48h、72h进行细胞计数以绘制细胞增殖曲线,转染后48h进行MTT实验以检测细胞增殖活力;免疫组织化学法检测cyclinD1和Ki67的表达,Western blotting法检测cyclinD1的表达情况。结果转染了靶向膜联蛋白A7的siRNA后48h的HepG2细胞,细胞计数和MTT实验可见siRNA干扰组细胞增殖活力较阴性对照组和空白对照组显著降低(P<0.05);cyclinD1和Ki67蛋白表达均为siRNA干扰组细胞表达显著低于两对照组(P<0.05)。结论膜联蛋白A7低表达可能对HepG2细胞的增殖具有一定的抑制作用。
Objective To investigate the effect of annexin A7 low expression on the proliferation of human hepatoma HepG2 cells. Methods Western blotting was used to identify siRNAs that could effectively inhibit the expression of annexin A7. Then siRNA was transfected into HepG2 cells by lipofection. The cells were divided into siRNA interference group, negative control group and blank control group. Cell counts were performed 24h, 48h, 72h after staining to determine the cell proliferation curve. MTT assay was performed 48h after transfection to detect cell proliferation activity. The expressions of cyclinD1 and Ki67 were detected by immunohistochemistry. The expression of cyclinD1 was detected by Western blotting. Results After HepG2 cells transfected with siRNA targeting Annexin A7 for 48h, the cell proliferation and viability of siRNA interference group were significantly decreased compared with those of negative control and blank control (P <0.05) The expression of all the siRNA interference group cells was significantly lower than the two control groups (P <0.05). Conclusion The low expression of annexin A7 may have some inhibitory effects on the proliferation of HepG2 cells.