甲胎蛋白调控HIPPO信号通路对肝细胞癌细胞重编程基因表达的影响OA

Objective To explore the effect of alpha-fetoprotein(AFP)regulating the HIPPO signaling pathway on the expression of reprogramming genes in hepatocellular carcinoma(HCC)cells.Methods Construct the HUH7 cell line with stable down-regulation of AFP and the HLE cell line with stable up-regulation of AFP According to the different transfected viruses,they were divided into the HUH7 group(untransfected),the HUH7C group(transfected with LV-NC),the HUH7-SIAFP group(transfected with LV-siAFP),HLE(untransfected),HLEC(transfected with LV-NC),and HLE-AFP(transfected with LV-AFP).The effect of AFP on the expression of reprogramming genes was detected by western bloting.The protein interaction between AFP and YAP was simulated by Discovery studio software.The correlation between YAP and reprogramming genes was analyzed by GEPIA2 online database.The effect of down-regulation or up-regulation of AFP on the colony formation of HCC cells was detected by plate cloning.The effect of down-regulation or up-regulation of AFP on the distribution of YAP protein in the cytoplasm and nucleus of HCC cells was observed by immunofluorescence assay.The interaction between AFP and YAP protein in HUH7 cells was detected by CO-IP assay.Results Upregulation of AFP can promote the expression of reprogramming genes in HCC cells,while downregulation of AFP can inhibit the expression of reprogramming genes in HCC cells.GEPIA2 online database analysis showed a positive correlation between YAP and reprogramming gene table.Discovery studio software simulation,CO-IP experiment,and immunofluorescence experiment suggested that AFP and YAP interact,and AFP can affect the nuclear localization of YAP.Conclusion In HCC cells,AFP may enhance the expression of downstream reprogramming genes by promoting the aggregation of YAP in the HIPPO signaling pathway in the nucleus.