ZNRF3 Promotes Apoptosis Of Hepatocellular Carcinoma Cells Through Hedgehog Signaling Pathway

Objective: This topic was research the expression of ZNRF3 in liver cancer tissues and adjacent tissues of liver cancer,then the fuction and mechanism of ZNRF3 on apoptosis of liver cancer cell Hep G2 was detected by measured with flow cytometry and Western Blot under ZNRF3 overexpression;Clarify ZNRF3 whether regulating the mechanism in the development of cancer of the liver,also provide a new marker for for the early detection and prognosis of liver cancer.Methods: Immunohistochemical detection the expression of ZNRF3 in hepatocellular carcinoma HCC tissues(40 cases)and adjacent tissues(30 cases).The overexpression effect of ZNRF3 in hepatocellular carcinoma cell line Hep G2 was detected by Western Blot;Moreover,The expriment were divided into three groups: I: control group;II: transfected with empty plasmid transfected group;III: overexpression of ZNRF3 plasmid to construct the recombinant plasmid of ZNRF3 group.By using the transient transfection,the plasmid and empty plasmid vectors were transfected into Hep G2 cells.The transfection efficiency was measured by fluorescence microscopy,flow cytometry was used to detect the apoptosis rate of Hep G2 cell;RT-PCR detection m RNA level of Smo andGli1,which were the the key component in the Hedgehog signaling pathway;Western blot detection the protein level of Smo and Gli1.Results: ZNRF3 was downregulated in liver cancer tissues compared to liver tissues.The results of transient transfection showed that Hep G2 cells observed in the view of most of the visible green fluorescence under the fluorescence microscope,the transfection efficiency is high(more than 80%).Flow cytometry results showed that: compared with I group and II group,the apoptosis rate of Hep G2 cells III group was significantly increased(P <0.05).The apoptosis rate of I group compared with II group have no significant difference(P = 0.05).Western-Blot detection the protein level of Smo and Gli1 results showed that compared with I group and II group,the protein expression of Smo and Gli1 in group III cells was significantly reduced(P<0.05),and There is no difference in the I group and II group(P = 0.05).The m RNA expression of Smo and Gli1 detected by RT-PCR assay revealed that compared with I group and II group,the m RNA levels of Smo and Gli1 in III group were significantly reduced(P<0.05),I group and II group compared with no significant difference(P = 0.05).Conclusions:ZNRF3 was downregulated in liver cancer tissues compared to liver tissues.And overexpression of ZNRF3 inhibited the Hedgehog /Smo /Gli1 signaling pathway to promoted proliferation of liver cancer cells.