The Effect Of AFP On HepG2 Migration And The Relationship Between AFP And KLF6 In Hepatocellular Carcinoma

Objective: To study the expression of AFP and KLF6 in hepatocellular carcinoma(HCC)tissues and HCC cell line.To investigate the effect of AFP on the migration of human HCC cell line HepG2 and to further explore the relationship between AFP and KLF6.Methods: A total of 30 specimens of fresh HCC tissues and paraffin-embedded specimens with clear pathological diagnosis and clinical data were collected from the Department of General Surgery,The First Hospital of Lanzhou University from Dec.2016 to Dec.2017.Among them,20 patients had high serum AFP(AFP>1000 IU/ml)and 10 patients had low serum AFP(AFP<15 IU/ml).Immunohistochemistry was used to detect the expression of AFP in HCC tissues.The expression level of KLF6 mRNA in 30 cases of hepatocellular carcinoma and matched paracancerous tissues were detected by RT-qPCR.The lentiviral vectors mediated recombinant AFP and AFP-shRNA were transferred into human HCC cell line HepG2 and the infection efficiency was detected by fluorescence inverted microscope.The overexpression and silence efficiency of AFP and the variation of KLF6 level was evaluated via RT-qPCR and Western Blot respectively.Wound healing assays and transwell assays were used to analyze the migration of human HepG2 cells.Results: Twenty cases of high serum AFP level showed a higher expression of AFP in HCC tissues,inversely,10 cases of low serum AFP showed low expression of AFP in HCC tissues.The KLF6 expression in cancer tissue was lower than that in paracancerous tissues.KLF6 expression is significantly lower in high serum AFP HCC patients than that in low serum AFP patients(P<0.01).The human HCC cell line HepG2 was successfully transfected with lentivirus mediated recombinant AFP and AFP-shRNA.Silencing AFP significantly enhanced KLF6 expression in HepG2 cells in vitro(P<0.05),whereas AFP overexpression reversed this phenotype(P<0.01).Functionally data showed that silencing AFP reduced the migration potential ofHepG2 cells(P<0.05),whereas AFP overexpression increased the migration potential of HepG2 cells(P<0.05).Conclusion: Serum AFP level in HCC patients is consistent with HCC tissues.Overexpression of AFP could significantly inhibit the KLF6 level.KLF6 expression in HCC tissues is correlated with the level of AFP,the former might regulated by AFP.AFP could promote HCC cell migration.