半乳糖凝集素3低表达对肝癌细胞增殖的影响及作用机制

Effect and mechanism of low expression of galectin 3 on the proliferation of hepatocellular cancer cells

目的探讨半乳糖凝集素3(Gal-3)低表达对肝癌HepG2细胞增殖的影响及作用机制。方法将细胞分为siRNA干扰组(转染靶向Gal-3的siRNA)、阴性对照组(转染阴性siRNA)和空白对照组(仅加转染试剂)。转染48 h后,采用实时qPCR和Western blotting在mRNA和蛋白水平进行抑制效果鉴定。MTT法检测Gal-3表达抑制后对细胞增殖活力的影响;Western blotting检测各组细胞增殖细胞核抗原(PCNA)、蛋白激酶(AKT)、磷酸化AKT(p-AKT)、哺乳动物雷帕霉素靶蛋白(mTOR)及磷酸化mTOR(p-mTOR)蛋白的表达。结果Western blotting和实时qPCR检测结果显示,siRNA干扰组Gal-3表达较阴性对照组和空白对照组均显著降低(P<0.05)。与阴性对照组和空白对照组比较,siRNA干扰组细胞增殖活力,PCNA、p-AKT及p-mTOR表达明显下调(P<0.05);而AKT、mTOR表达无统计学差异(P>0.05)。结论Gal-3低表达抑制肝癌HepG2细胞增殖,其作用机制可能是通过抑制PI3K/AKT/mTOR信号转导通路来实现的。

Objective To investigate the effect and mechanism of low expression of galectin 3(Gal-3)on the proliferation of human hepatocellular cancer(HepG2)cells.Methods HepG2 cells were divided into siRNA interference,negative control,and blank control groups.Using liposome transfection,siRNA targeted to Gal-3 and negative siRNA were transfected into the siRNA interference and negative control groups,respectively.Transfection reagent was added in the blank control group.After 48 h of transfection,inhibitory effects were identified at the mRNA and protein levels with RT-qPCR and Western blotting,respectively.The MTT assay was used to detect the effect of low expression of Gal-3 on proliferation of HepG2 cells.Western blotting was used to detect the expression levels of proliferating cell nuclear antigen(PCNA),protein kinase(AKT),p-AKT,mammalian target of rapamcycin(mTOR),and p-mTOR.Results Western blotting and RT-qPCR results showed that the expression of Gal-3 in the siRNA interference group was significantly lower than that in the negative control and blank control groups(P<0.05).The proliferation activity and expression of PCNA,p-AKT,and p-mTOR of the siRNA interference group were significantly decreased compared to the negative control and blank control groups(P<0.05).However,there was no statistically significant difference in the expression of AKT and mTOR amongst these groups(P>0.05).Conclusion Low expression of Gal-3 prevents the proliferation of HepG2 cells by inhibiting the PI3K/AKT/mTOR signaling pathway.