m6A去甲基化酶ALKBH5在肝细胞癌发生发展中的功能作用OACSTPCD

Objective To explore the functional role of m6A demethylase ALKBH5 in hepatocellular carcinoma(HCC).Methods The mRNA and protein expression of ALKBH5 were analyzed in HCC and para-cancerous tissues using public databases(TCGA,CPTAC).The expression of ALKBH5 was also assessed in normal hepatocytes LO2 and HCC cell lines HepG2,Hep3B,and Huh7 through qRT-PCR.Stably knocked-down HCC cell lines were generated using siRNA interference and lentiviral techniques to test the interference efficiency at the level of ALKBH5 mRNA and protein.Subsequently,ALKBH5 expression in Hep3B and Huh7 was examined by CCK-8,plate cloning,cell scratching,and transwell tests to analyze the effect of ALKBH5 on the proliferative activity,migration,and invasion ability of HCC.Results In TCGA and CPTAC databases,ALKBH5 was highly expressed at both mRNA[(5.78±0.62)vs(5.41±0.25)]and protein[(0.09±0.35)vs(-0.01±0.16)]levels in HCC tissues compared to para-cancerous tissues.ALKBH5 expression in LO2(1.0±0.1)was significantly lower than that in Hep3B(1 430.7±103.9),Huh7(1 515.6±162.4)and HepG2(311.0±29.6).Hep3B and Huh7 with stable knockdown of ALKBH5 were successfully constructed by the lentiviral method.The proliferative activity,migration and invasion ability of Hep3B and Huh7 cells were significantly attenuated after inhibiting the expression of ALKBH5(P<0.05).Conclusion ALKBH5 exhibited high expression in HCC cells,and its downregulation led to a suppressive effect on the malignant biological behavior of HCC.Further investigations are needed to elucidate other biological functions of ALKBH5 in HCC and its associated regulatory network.