Effects Of Co-regulation Of P21 And MTH1 Gene Expression On The Growth Of Human Hepatoma Cell Line HepG2 Cells

Objective: Hepato cellular carcinoma(HCC)is one of the most common malignant cancers in terms of both incidence and mortality worldwide.Because of the unobservable cause and lack of effective diagnostic methods at the early stage,almost all the patients discover hepato cellular carcinoma at the late stage with the spread and metastasis of cancer cells.Up to now,there is still no satisfactory therapeutic strategy to increase the survival rate of patients.Thus,it is very important to understand the mechanisms of HCC and find a new therapy.This study aims to investigate the migration,invasion,proliferation and apoptosis of Hep G2 cells through up-regulating p21 gene and down-regulating MTH1 gene.Methods: The sa RNA targeting p21 promoter and the si RNA targeting MTH1 were transfected into Hep G2 cells by transfection reagent.According to the different transfection mixtures,cells were divided into five groups: CT group?NC group?p21 group?MTH1 group and p21+MTH1 group.q PCR was used to detect the expression of p21 and MTH1;Western Blot was used to detect the expression of p21,MTH1 and some apoptosis proteins.CCK8 assay was used to detect the cell proliferation;scratch assay was used to detect the cell migration;Transwell invasion assay was used to detect the cell invasion;flow cytometry was used to detect the cell apoptosis.Results: Hep G2 cells were transfected by sa RAN-p21 and si RNA-MTH1.The q PCR and Western Blot showed that the expression of p21 gene was significantly increased by sa RAN-p21(P<0.01)and the expression of MTH1 gene was significantly reduced by si RNA-MTH1(P<0.01).The Western Blot showed that the expression of Bak and Caspase-3 were significantly increased(P<0.05).Both the cell proliferation and the scratch healing were significantly decreased(P<0.05);the number of Hep G2 cells that passed through the basement membrane was significantly reduced(P<0.01)compared with CT group.CT group and NC group had no statistical significance(P>0.05).The proliferation,migration and invasion of the co-transfected cells were better inhibited than the sa RAN-p21 or si RAN-MTH1 transfected cells(P<0.05).Conclusion: The expression of p21 gene can be effectively upregulated by sa RNA-p21,and the expression of MTH1 gene can be effectively downregulated by si RNA-MTH1.Upregulating the expression of p21 gene and downregulating the expression of MTH1 gene can inhibit the migration,invasion and proliferation of Hep G2 cells.And it may enhance apoptosis via Bak/Bcl-xl signaling pathway.Co-regulation of p21 and MTH1 gene expression showed a more significant effect than only regulating a single gene.