The Role And Molecular Mechanism Of The MARCH1 In The Development And Progression Of Hepatocellular Carcinoma

Aim:To investigate the role and molecular mechanism of MARCH1 in the development and progression of hepatocellular carcinoma(HCC)Methods:Here,we determined the protein MARCH1 level in human hepatocellular carcinoma tissues,adjacent non-tumour liver tissues,HCC cells(HepG2,Hep3B),and nomal hepatocyte cells(HL-7702,HHL-5)by immunohistochemical(IHC)and western blot.Cell proliferation,apoptosis,migration,and invasion analysis were performed in the HepG2 and Hep3B cells treated with MARCH1 siRNA,MARCH1 overexpressed plasmid,pirarubicin(THP)and secalonic acid-F(SAF);And western blot analysis was used to detect the protein level of MARCH1,PI3K,AKT,p-AKT,?-catenin,Mcl-1,Bcl-2,cleaved caspase-3,cleaved caspase-7 in the treated HepG2 and Hep3B cells.In addition,the model of HCC transplantation in nude mice was established.the growth of hepatocellular carcinoma were recorded in nude mice,which were given an intra-tumor injection of siRNA targeting MARCH1,or were intragastrically(Ig)administered SAF.Additionally,all nude mice HCC xenograf model were subjected to magnetic resonance imaging(MRI);the apparent diffusion coefficient(ADC)values,derived from diffusion-weighted imaging(DWI),were measured in tumors.Moreover,Western blot analysis was used to detect the protein level of MARCH1 and PI3K,AKT,p-AKT,?-catenin,Mcl-1,Bcl-2,cleaved caspase-3,cleaved caspase-7 of PI3K-AKT-?-catenin pathways in tumors.Results:MARCH1 was obviously highly expressed in human hepatocellular carcinoma tissues and cells(HepG2,Hep3B)compared to adjacent non-tumour liver tissues and nomal hepatocyte cells(HL-7702,HHL-5).Our findings also show that the proliferation,migration,and invasion of hepatocellular carcinoma were suppressed,but the apoptosis was increased,as a result of MARCH1 knockdown by either siRNA targeting MARCH1,THP treatment or SAF treatment.Conversely,the proliferation,migration,and invasion of hepatocellular carcinoma was obviously accelerated by transfecting the MARCH1 plasmid to make MARCH1 overexpressed.MARCH1 downregulation could downregulate the protein level of PI3K,p-AKT,?-catenin,Mcl-1 and Bcl-2,while upregulation of the cleaved caspase-3,cleaved caspase-7;Conversely,MARCH1 overexpressed could upregulate the protein level of PI3K,p-AKT,?-catenin,Mcl-1 and Bcl-2.Moreover,in vivo,the results exhibited a significant inhibition of the growth of hepatocellular carcinoma of the reduced tumor volume and tumor weight,and increased ADC values in tumors of nude mice,which were given an intra-tumor injection of siRNA targeting MARCH1 or SAF treatment.Western blot showed that the protein level of MARCH1,PI3K,p-AKT,?-catenin,Mcl-1 and Bcl-2 were decreased,while the cleaved caspase-3 and cleaved caspase-7 were increased in tumors treated with MARCH 1 siRNA or SAF.Our results confirmed that MARCH1 siRNA and SAF could inhibit the progression of hepatocellular carcinoma by downregulating the MARCH1 and PI3K/AKT/?-catenin signaling pathway.Conclusion:MARCH1 was aberrantly and highly expressed in HCC;and MARCH 1 has an important role in the development and progression of hepatocellular carcinoma by regulating the PI3K/AKT/?-catenin signaling pathways.Therefore,MARCH1 is a potential molecular target for HCC treatment,and SAF is a promising agent targeting MARCH1 to treat liver cancer patients.