Preliminary Study On Role Of ZNF439 In The Proliferation And Metastasis Of Hepatocellular Carcinoma

ObjectiveThis research aimed to investigate the expression of zinc finger protein 439(ZNF439)in hepatocellular carcinoma and para-carcinoma liver tissues,and preliminarily investigated its role in the development of hepatocellular carcinoma.And overexpressed ZNF439 in hepatocellular carcinoma cell to further studied the role of ZNF439 in tumor proliferation and metastasis of hepatocellular carcinoma.MethodsThe expression of ZNF439 was examined in 21 paired hepatocellular carcinoma tissues and their para-carcinoma liver tissues by immunohistochemistry(IHC).The expression of ZNF439 in hepatocellular carcinoma tissues and their para-carcinoma liver tissues were analyzed and compared.Then,we generated two ZNF439 over-expression hepatocellular carcinoma cell lines by G418/Geneticin positive selection,including Hep-3b and LM3.In these two cell lines,cell proliferation test was performed to investigate the role of ZNF439 in tumor proliferation of hepatocellular carcinoma by MTS assay.In addition,we had applied cell migration test,stained the cell cytoskeleton with Rhodamine-Phalloidin and tube formation assay to investigate the role of ZNF439 in tumor metastasis of hepatocellular carcinoma.ResultsImmunohistochemistry(IHC)showed ZNF439 expressed in nucles only.In the nucleus of hepatocellular carcinoma cells,ZNF439 mainly showed negative expression and weak positive expression,accounting for 66.7%and 28.6%of the total cases respectively,followed by the moderate positive expression,only accounting for 4.7%of the total cases.While ZNF439 showed strong expression in the most cell nucleus of para-carcinoma liver tissues,including 38.1%cases of strong positive expression,28.6%cases of moderate positive expression and followed by weak positive expression,accounting for 33.3%of the total cases.There was significant difference between the two groups(P<0.0001).We Successfully generated ZNF439 over-expression stable cell lines in Hep-3b and LM3,which were verified by Semi-RT PCR and Western Blot.MTS assay showed that ZNF439 over-expression had any influence on the proliferation in both Hep-3b and LM3 cell lines.Cell migration assay demonstrated that tumor migration ability was markedly decreased in the condition of ZNF439 over-expression in both Hep-3b and LM3 cell lines.And it was found that the cells cytoskeleton of ZNF439 over-expression cells was reorganized less compared to the Hep3b-MOCK and LM3-MOCK cells.The result of tube formation assay had showed LM3-ZNF439 cells inhibited ability of tube formation.However,no angiogenesis was observed in Hep-3b cells.ConclusionThis research showed significantly differential distribution of ZNF439 between liver tumor tissues and para-carcinoma liver tissues,demonstrating ZNF439 may play a role in the development of hepatocellular carcinoma.ZNF439 does not affect the proliferation ability of hepatocellular carcinoma.While ZNF439 could suppress tumor metastasis and reorganize tumor cell cytoskeleton in hepatocellular carcinoma.ZNF439 may restrain cytoskeleton reorganization and inhibit vasculogenic mimicry of hepatocellular carcinoma cell to metastasis.