HSP70对HepG22.15细胞HBV表达及Notch1信号通路的影响

Effects of HSP70 on HBV expression and Notch1 signaling pathway in HepG22.15 cells

目的研究热休克蛋白70(HSP70)对HepG22.15细胞HBV表达及Notch1信号通路的影响。方法实验分组:空白对照组(HepG22.15细胞不做任何处理);空载组:(转染pENTER空载质粒);HSP70过表达组(转染HSP70过表达质粒)。比较3组的HBsAg、HBeAg表达及HBV DNA载量,探讨HSP70对HepG22.15细胞HBV表达的影响,采用Western blot检测Notch1和Hesl蛋白表达,探讨HSP70对HepG22.15细胞Notch1信号通路的影响。结果与空白对照组相比,空载组HBsAg、HBeAg阳性表达差异无统计学意义(P>0.05);与空载组相比,HSP70过表达组HBsAg、HBeAg阳性表达显著增加(P<0.05)。与空白对照组相比,空载组HBV-DNA浓度无改变(P>0.05);与空载组相比,HSP70过表达后,细胞上清液中HBV-DNA水平显著升高(P<0.05)。与空白对照组相比,Hes1、Notch1蛋白浓度在HSP70过表达组显著升高(P<0.05);与空载组相比,Hes1、Notch1蛋白浓度在HSP70过表达组显著升高(P<0.05)。结论HSP70过表达促进HBV复制,并且上调Notch1、Hes1蛋白水平,HSP70可能通过促进HBV表达和激活Notch1信号通路影响肝癌的发生发展及预后。

Objective To investigate the effects of HSP70 on HBV expression and Notch1 signaling pathway in HepG22.15 cells.Methods The experiment was divided into thre groups,the blank control group(the HepG22.15 cells without ant treatment),vector group(transfected by pENTER plasmid),and HSP70 overexpression group(transfected with HSP70 overexpression plasmid).The expression levels of HBsAg and HBeAg and HBV DNA load were detected and compared among the three groups to investigate the effects of HSP70 on HBV expression in HepG22.15 cells.Moreover Western Blot was used to detect the expression levels of Notch1 and Hesl protein to analyze the effects of HSP70 on Notch1 signaling pathway.Results There were no significant differences in the expression levels of HBsAg and HBeAg between vector group and blank control group(P>0.05).As compared with those in vector group,the expression levels of HBsAg and HBeAg in HSP70 overexpression group were increased significantly.As compared with those in blank control group,there was no change in HBV-DNA concentration in vector group(P>0.05).As compared with those in vector group,the expression levels of HBV-DNA in cell supernatant were increased significantly after the overexpression of HSP70.As compared with those in blank control group,the levels of Hes1 and Notch1 protein were significantly increased in the HSP70 overexpression group,moreover,which in HSP70 overexpression group were significantly increased,as compared with those in vector group(P<0.05).Conclusion HSP70 overexpression can promote HBV replication and up-regulate the expression levels of Notch1 and Hes1 protein.HSP70 may affect the occurrence,development and prognosis of hepatocellular carcinoma by promoting HBV expression and activating Notch1 signaling pathway.