MiR-142-3p Inhibits Aerobic Glycolysis And Cell Proliferation In Hepatocellular Carcinoma Via Targeting LDHA

Objective1 To screen of low expression and metabolism related miRNA in liver cancer:mir-142-3p;2 To identify the role of miR-142-3p in regulation of hepatocellular carcinoma aerobic glycolysis and proliferation.3 To find out the molecular mechanism of these roles.Contents and methods1.The low expression of mir-142-3p in liver cancer has significant correlation with energy metabolism.1)The low-expression miRNAs through bioinformatics analysis of TCGA database and GEO data set GSE 22058,GSE 54751,GSE 21362 and GSE 12717 contains:hsa-miR-424;hsa-miR-195;hsa-let-7c;hsa-miR-130a;hsa-miR-142-3p;hsa-miR-29c;hsa-miR-10a;hsa-miR-99a;hsa-miR-378?2)miR-142-3p and miR-195 were significantly associated with energy metabolism through the biological function of pathway analysis.2.miR-142-3p suppresses aerobic glycolysis and proliferation of hepatoma carcinoma cell.1)Use of miR-142-3p mimic transfection to hepatocellular carcinoma cell line SK-Hep-1 and Huh-7 to overexpress miR-142-3p;2)To detect the consumption of glucose and production of lactate in medium after overexpressing miR-142-3p.3)Use of EDU and CCK 8 experiments to detect cell proliferation ability after expressing miR-142-3p.3 The molecular mechanism of miR-142-3p controlling aerobic glycolysis and proliferation.1)Using RNA22(http://cbcsrv.watson.ibm.com/rna22.html)to predict miR-142-3p target genes.2)Using Western blot to dectect if the protein level of LDHA is influenced by miR-142-3p.3)Using the dual luciferase experiment to identify the direct target relationship between LDHA and miR-142-3p;4)To prove that miR-142-3p regulates aerobic glycolysis and proliferation by LDHA through the RESCUE experiment.Results1.The low expression of miR-142-3p in liver cancer has significant correlation with energy metabolism.1)Through the analysis of bioinformatics,miRNAs in TCGA database and GEO data sets GSE 22058,GSE 54751,GSE 21362 and GSE 12717 which were down-regulated included:hsa-mir-424;Hsa-miR-195;Hsa-let-7 c;Hsa-miR-130a;Hsa-miR-142-3p;Hsa-miR-29c;Hsa-miR-10a;Hsa-miR-99-a;Hsa-miR-378.2)miR-142-3p and miR-195 were significantly correlated with energy metabolism by analyzing the biological function pathway of each miRNA target genes.2.miR-142-3p inhibited the aerobic glycolysis and proliferation of hepatocellular carcinoma cells.1)The glycolysis activity of hepatocellular carcinoma cell lines sk-hep-1 and huh-7 was weakened after the overexpression of miR-142-3p compared with the scramble.2)Compared with the scramble,the proliferation of hepatocellular carcinoma cells was inhibited after overexpressing mir-142-3p.3.miR-142-3p directly targeted LDHA to lower its expression,thereby inhibiting the aerobic glycolysis and proliferation of hepatocellular carcinoma cells.1)miR-142-3p can lower the protein level of LDHA,and the double luciferase experiment indicates that mir-142-3p can directly target LDHA;2)Overexpressing the expression of mir-142-3p and overexpressing LDHA at the same time,to detect the aerobic glycolysis and proliferation of hepatocellular carcinoma cells,can play a rescue role.ConclusionmiR-142-3p is low expression in liver cancer tissues and has significant correlation with energy metabolism.Overexpression of miR-142-3p can inhibit the aerobic glycolysis and proliferation of hepatocellular carcinoma cells.On the mechanism,miR-142-3p directly targeted LDHA to lower its expression,thereby inhibiting the aerobic glycolysis and proliferation of hepatocellular carcinoma cells.