ACTR调控IGF-1R对肝癌细胞生长的影响

Regulatory effect of ACTR on the proliferation of hepatocellular carcinoma cells by mediating IGF-1R and the clinical significance

目的探究ACTR调控IGF-1R的表达对肝癌细胞生长的影响及意义。方法外源性免疫共沉淀、内源性免疫共沉淀检测ACTR与IGF-1R之间的相互作用。应用细胞免疫荧光共定位,检测ACTR与IGF-1R在肝癌细胞内共定位情况。qRT-PCR及Western blot实验,检测肝癌细胞中ACTR对IGF-1R表达的影响。将肝癌细胞系分为阴性对照组、转染ACTR组、敲低IGF-1R组、敲低IGF-1R+转染ACTR组,应用CCK8法测定各组细胞生长情况。结果外源性免疫共沉淀、内源性免疫共沉淀均证实ACTR与IGF-1R之间存在相互作用。细胞免疫荧光共定位证实ACTR与IGF-1R在肝癌细胞内存在共定位表达。qRT-PCR证实敲低ACTR后,IGF-1R mRNA表达水平降低(P<0.05),回转ACTR后IGF-1R mRNA表达水平亦可恢复。Western blot证实敲低ACTR后IGF-1R表达降低,而回转ACTR后IGF-1R表达可恢复。细胞生长曲线说明ACTR能够促进肝癌细胞的生长(P<0.05),当应用siRNA敲低IGF-1R后,ACTR的促细胞生长作用减弱。结论ACTR与IGF-1R之间存在相互的作用,IGF-1R介导了ACTR促进肝癌细胞生长的过程,ACTR/IGF-1R轴在肝癌细胞的生长中发挥了必要的作用,有可能成为治疗肝癌的靶点之一,为肝癌的防治提供新的思路。

Objective To investigate the regulatory effect of antibody-coupled T cell receptor(ACTR)on the proliferation of hepatocellular carcinoma cells by mediating insulin-like growth factor type 1 receptor(IGF-1R)and the clinical significance.Methods The interaction between ACTR and IGF-1R was detected by exogenous and endogenous coimmunoprecipitation.Immunofluorescence was used to detect the co-localization of ACTR and IGF-1R in hepatocellular carcinoma cells.The regulatory effect of ACTR on the mRNA and protein levels of IGF-1R was detected by Quantitative real-time polymerase chain reaction(QRT-PCR)and Western blot,respectively.The hepatocellular carcinoma cell line HepG2 was transfected with corresponding plasmids to overexpress or knock down of ACTR and IGF-1R,followed by measurement of cell growth via cell counting kit-8(CCK-8)assays.Results Both exogenous and endogenous coimmunoprecipitation confirmed the interaction between ACTR and IGF-1R.The co-localization of ACTR and IGF-1R in HCC cells was confirmed by immunofluorescence.Knockdown of ACTR significantly downregulated the mRNA and protein levels of IGF-1R,which were significantly reversed by overexpression of ACTR(P<0.05).The cellular growth curve showed that overexpression of ACTR promoted the growth of HepG2 cells,and its promotive effect on cell proliferation was weakened by knockdown of IGF-1R(P<0.05).Conclusion ACTR interacts with IGF-1R,which is involved in the process of ACTR in promoting the proliferation of HCC cells.The ACTR/IGF-1R axis plays a important role in the growth of HCC cells,which may become promising targets for the treatment of hepatocellular cancer.Our findings provide new ideas for the prevention and treatment of hepatocellular cancer.