MiR-21-5p Promotes Aerobic Glycolysis And Cell Proliferation In Hepatocellular Carcinoma By Targeting PFKM

Background and Purpose:HCC as one of the most common malignant tumors in the world,is still a serious threat to human health.The prognosis and long-term survival of patients with hepatocellular carcinoma are still poor because the diagnosis and treatment protocol commonly used in clinic is still difficult to make accurate early diagnosis of hepatocellular carcinoma.Micrornas(mirnas)are a class of short non-coding single-stranded RNA molecules encoded by endogenous genes with a length of about 20-24 nucleotides.It regulates the expression of target genes by cutting off the m RNA molecules of target genes and inhibiting the translation of target genes,that is,it regulates gene expression at the post-transcriptional level.Micro RNA-21-5p(mi R-21-5p),as one of the most widely recognized and highly conserved mirnas,has been well studied,but its effects on HCC and its mechanism of action remain unclear.The main purpose of this study is to explore the role and mechanism of mi R-21-5p in hepatocellular carcinoma.Methods:1.By searching GEO and TCGA databases,the overexpressed mirnas in hepatocellular carcinoma were screened.After further screening,the study object was selected as mi R-21-5p,and its target gene was screened as PFKM by target gene prediction software Target Scan.The targeting relationship between mi RNA-21-5p and the target gene PFKM was verified by double luciferase assay.2.Western blot and q RT-PCR were used to investigate the expression of mi R-21-5p and its target gene in liver cancer cells.3.Overexpression of mi RNA-21-5p and control liver cancer cells were constructed by cell transfection,and the effects of mi RNA-21-5p on glucose metabolism of liver cancer cells were investigated by lactic acid generation experiment and glucose consumption experiment.4.The effects of mi RNA-21-5p and its target genes on the proliferation of hepatoma cells were detected by CCK-8 assay.Results:1.It was verified that mi RNA-21-5p was overexpressed in hepatocellular carcinoma by searching relevant databases and q RT-PCR.2.The results of glucose consumption experiment and lactic acid production experiment indicate that mi RNA-21-5p overexpression group can promote glucose consumption and lactic acid production of liver cancer cells compared with the control group,that is,promote aerobic glycolysis of liver cancer cells.3.CCK-8 experiment proved that mi RNA-21-5p overexpression could promote the proliferation of hepatoma cells.4.The results of Western blot,immunohistochemistry and qRT-PCR showed a positive correlation between PFKM expression and mi RNA expression in liver cancer cells.The results of double luciferase assay indicated that PFKM was the target gene of mi RNA-21-5p.Conclusion:The experimental results of this study showed that mi RNA-21-5p regulates aerobic glycolysis of hepatocellular carcinoma by targeting PFKM to further promote the proliferation of hepatocellular carcinoma.