SPARCL1基因对肝癌SMMC-7721细胞增殖与凋亡的影响

［摘要］目的: 探讨富含半胱氨酸酸性分泌型糖蛋白类似物1(secreted protein acidic and rich in cysteines like 1，SPARCL1)基因对肝癌SMMC 7721细胞增殖与凋亡的影响。方法: 将肝癌SMMC 7721细胞随机分为5组，即空白对照组，pIRES2 ZsGreen组，pIRES2 ZsGreen SPARCL1组，阴性 siRNA组和SPARCL1 siRNA组，空白对照组不转染，其余4组分别转染pIRES2 ZsGreen空质粒、pIRES2 ZsGreen SPARCL1、阴性 siRNA及SPARCL1 siRNA；蛋白质印迹法检测SPARCL1蛋白表达，同时联合荧光显微镜检测转染效果；MTT法检测各组细胞增殖能力；流式细胞术检测各组细胞凋亡情况。 结果： 蛋白质印迹和荧光镜结果同时证实转染效果良好。MTT结果显示5组细胞抑制率差异有统计学意义(F=55.45，P＜0.01)，pIRES2 ZsGreen SPARCL1组增殖抑制率明显高于其余4组(P＜0.05)；SPARCL1 siRNA组细胞增殖抑制率低于其余4组(P＜0.05)。流式细胞结果显示pIRES2 ZsGreen SPARCL1组凋亡率明显高于其余4组 (P<0.05)；SPARCL1 siRNA组凋亡率明显低于其余4组(P＜0.05)。结论： 过表达SPARCL1抑制肝癌细胞SMMC 7721生长和促进凋亡，沉默SPARCL1则反之。

Influence of SPARCL1 gene on the proliferation and apoptosis of human hepatocellular carcinoma SMMC-7721 cell

［Abstract］Objective To investigate the influence of secreted protein acidic and rich in cysteines like 1（SPARCL1） gene on the proliferation and apoptosis of human hepatocellular carcinoma SMMC 7721 cell. Methods SMMC 7721 cells were divided into five groups, including blank control group, pIRES2 ZsGreen vector group, pIRES2 ZsGreen SPARCL1 group, negative siRNA group and SPARCL1 siRNA group. Western blotting was used to investigate SPARCL1 expression, while fluorescence microscope were used to verify the transfection efficiency of SPARCL1. MTT assay was used to detect cell proliferation ability; and flow cytometry was used to measure apoptosis rate. Results The transfection efficiency of SPARCL1 was high through the fluorescence microscope observation and western blotting. SPARCL1 expression, the cell inhibition ratio, and the apoptosis ratio in pIRES2 ZsGreen SPARCL1 group was significantly higher than other four groups(P<0.05), while SPARCL1 siRNA group was lower than other four groups(P<0.05). Conclusion SPARCL1 overexpression elevated the proliferation inhibition ratio and apoptosis ratio of human hepatocellular carcinoma SMMC 7721 cell； on the contrary， SPARCL1 silencing lowered.