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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,p IRES2-Zs Green组,pIRES2-Zs Green-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p IRES2-Zs Green空质粒、pIRES2-Zs Green-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),p IRES2-Zs Green-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-Zs Green-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。
Objective: To investigate the effects of secreted protein acidic and rich in cysteines-like 1 (SPARCL1) gene on the proliferation and apoptosis of hepatoma SMMC-7721 cells. Methods: The hepatocellular carcinoma SMMC-7721 cells were randomly divided into 5 groups: blank control group, pIRES2-Zs Green group, pIRES2-Zs Green-SPARCL1 group, negative siRNA group and SPARCL1 siRNA group. Four groups were transfected with pIRES2-Zs Green empty plasmid, pIRES2-Zs Green-SPARCL1, negative siRNA and SPARCL1 siRNA respectively. Western blotting was used to detect the expression of SPARCL1 protein and fluorescence microscopy. MTT assay was used to detect cell proliferation Ability; flow cytometry detection of apoptosis in each group. Results: Western blotting and fluoroscopic results confirmed that the transfection effect was good. The MTT results showed that there was significant difference between the 5 groups (F = 55.45, P <0.01), the inhibition rate of pESES-Zs Green-SPARCL1 group was significantly higher than the other 4 groups The proliferation inhibition rate was lower than the other 4 groups (P <0.05). The results of flow cytometry showed that the apoptosis rate of pIRES2-Zs Green-SPARCL1 group was significantly higher than that of the other 4 groups (P <0.05). The apoptosis rate of SPARCL1 siRNA group was significantly lower than the other 4 groups (P <0.05). Conclusion: Overexpression of SPARCL1 can inhibit the growth and promote the apoptosis of hepatocellular carcinoma SMMC-7721, whereas the silencing of SPARCL1 conversely.