POU5F1B Promotes Hepatocellularcarcinoma Proliferation By Activating AKT And MiR-660 Promotes HCC Cellproliferation By Targeting PPP2R2A

Objective: To detect the expression of POU5F1 B in hepatocellular carcinoma(HCC)and explore its regulatory mechanism in promoting the proliferation of hepatocellular carcinoma.Methods: The TCGA database was used to analyze the association of POU5F1 B with the development of hepatocellular carcinoma.MTT assay,soft agar growth assay,Brd U incorporation assay and cell cycle assay were used to detect the effect of POU5F1 B overexpression on the proliferation of hepatocellular carcinoma cells and its regulation of AKT.Collecting the liver tissue of hepatocellular carcinoma patients and normal liver tissues from the affiliated hospital of Soochow University,and to detect the relationship between expression of POU5F1 B and the expression levels of Cyclin D1,p21 and p27,and the phosphorylation of AKT in tissues by Western blotting and real-time PCR.Results: TCGA database analysis showed that POU5F1 B was significantly up-regulated in hepatocellular carcinoma cells and tissues.Liver cancer patients with high expression of POU5F1 B had shorter life span than those with low expression of POU5F1 B.In MTT assay,soft agar growth assay,Brd U incorporation assay and cell cycle assay,it was found that overexpression of POU5F1 B can promote the proliferation of hepatoma cells,and knocking down POU5F1 B inhibits the proliferation of hepatoma cells,and POU5F1 B can activate AKT,AKT inhibition in HCC cells with POU5F1 B overexpression significantly inhibited cell proliferation compared to cells only with POU5F1 B overexpression,indicating that POU5F1 B promotes the proliferation of hepatoma cells by activating AKT.The detection of samples obtained from HCC showed that the expression level of POU5F1 B was positively correlated with the expression level of Cyclin D1,but negatively correlated with the expression levels of p21 and p27 and the phosphorylation level of AKT.Conclusion: POU5F1 B promotes the proliferation of hepatocellular carcinoma by activating AKT,and provides a new target for the treatment of liver cancer.Objective: To detect the expression of miR-660 in hepatocellular carcinoma(HCC)and explore its regulatory mechanism in promoting the proliferation of hepatocellular carcinoma.Methods: The TCGA database was used to analyze the association of miR-660 with the development of hepatocellular carcinoma.The expression of miR-660 in various liver cancer cells and tissues was detected by real-time PCR assay.MTT assay,soft agar growth assay,Brd U incorporation assay and cell cycle assay were used to detect the effect of miR-660 overexpression on the proliferation of hepatoma cells and the regulation of PPP2A2 R.The regulation of PPP2A2 R by miR-660 was detected by luciferase reporter gene assay,real-time PCR assay and Western blotting.The relationship between miR-660 and the expression of CCND1 and p21 was detected by real-time PCR.Results: The TCGA database analysis showed that miR-660 was significantly up-regulated in hepatoma cells and tissues.Real-time PCR experiments showed that miR-660 was upregulated in multiple hepatoma cells.In MTT assay,soft agar growth assay,Brd U incorporation assay and cell cycle assay,it was found that overexpression of miR-660 can promote the proliferation of hepatocellular carcinoma cells,and knocking down miR-660 inhibits the proliferation of hepatoma cells.And miR-660 can inhibit the level of PPP2R2 A,luciferase assay detected the direct binding of miR-660 and PPP2R2 A 3'-UTR.Real-time PCR experiments found that miR-660 inhibits p21 expression and promotes CCND1 expression.The soft agar growth experiment and Brd U incorporation experiment proved that knocking down PPP2R2 A in miR-660 knocking down Hep G2 cells can promote Hep G2 proliferation.Conclusion: miR-660 promotes the proliferation of hepatocellular carcinoma by targeting PPP2R2 A,and provides a new target for the treatment of hepatocellular carcinoma.