Expression And Function Of CCNA2 And CCNB1 Gene In Hepatocellular Carcinoma

Objective:The purpose of this study is to explore the expression levels of cyclin A2 and cyclin B1(CCNA2,CCNB1)in hepatocellular carcinoma and their effects on the proliferation and prognosis of hepatocellular carcinoma cells,and to explore their possible functions,so as to provide new ideas for the diagnosis and treatment of hepatocellular carcinoma.Method:1.Bioinformatics method was used to analyze the expression of CCNA2 and CCNB1 in liver cancer tissues by SMD forest map.2.The expression of CCNA2 and CCNB1 in liver cancer tissues and adjacent tissues in tissue microarray was determined by immunohistochemical assay.3.Human liver cancer cell lines Huh7 and SK-Hep-1 were collected as experimental materials.The expressions of CCNA2 and CCNB1 in Huh7 cells and SK-Hep-1 were down-regulated and up-regulated by lentiviral vector transfection technique,and the transfection efficiency was verified by real-time quantitative fluorescence PCR(q PCR)technique,and stable cell lines were obtained.4.The relationship between down-regulated and up-regulated CCNA2 and CCNB1 genes and cell proliferation was detected by CCK-8 proliferation assay.5.Kaplan Meier-Plotter was used to analyze the relationship between CCNA2,CCNB1 and 5-year survival rate in patients with different groups of hepatocytes.Result:1.The SMD forest map showed that the expression of CCNA2 in HCC was significantly higher than that in non cancer control liver tissue,and the random effect model showed that the combined SMD was 1.58(95%CI:1.42-1.75).I~2 was 78.0%(P<0.01);the expression of CCNB1 in HCC was higher than that in non cancerous tissues,and the combined SMD was 2.04(95%CI:1.82-2.27).I~2 was 76.0%(P<0.01).2.Immunohistochemistry results showed that the expression of CCNA2 and CCNB1 in HCC tissues were higher than that in adjacent tissues,the difference was statistically significant(P<0.0001).3.The results of q PCR showed that compared with NC group,CCNA2m RNA expression level of SK-Hep-1 hepatoma cells in Si group was down regulated(P<0.001);CCNA2 m RNA expression level of SK-Hep-1 hepatoma cells in OE group was up-regulated(P<0.01);CCNA2 m RNA expression level of Huh7 hepatoma cells in Si group was down regulated(P<0.001);The m RNA expression of CCNA2 was up-regulated in the OE group(P<0.01).Compared with the corresponding NC group,CCNB1 m RNA expression of SK-Hep-1hepatoma cells in Si group was down regulated(P<0.001);CCNB1 m RNA expression of SK-Hep-1 hepatoma cells in OE group was up-regulated(P<0.05);CCNB1 m RNA expression of Huh7 hepatoma cells in Si group was down regulated(P<0.001);CCNB1 m RNA expression of Huh7 hepatoma cells in OE group was up-regulated(P<0.01).4.CCK-8 proliferation experiment results showed that:after up regulating CCNB1 expression,the proliferation ability of Huh7 cells in OE group was significantly higher than that in NC-OE group(P<0.05);after down regulating CCNB1 expression,the proliferation ability of Huh7 cells in Si group was lower than that in NC-Si Group(P<0.05).5.Kaplan Meier plotter analysis showed that the overall survival rate of patients with high expression of CCNA2 and CCNB1 was lower than that of patients with low expression of CCNA2 and CCNB1;the overall survival rate of patients with high expression of CCNA2 and CCNB1 was lower than that of patients with low expression of CCNA2 and CCNB1 in patients with drinking history,no drinking history and no history of hepatitis virus infection(P<0.05).Conclusion:1.CCNA2 and CCNB1 were highly expressed in both HCC tissues and HCC cells.2.Up regulation of CCNB1 can promote the proliferation of Huh7 cells,down regulation of CCNB1 can inhibit the proliferation of Huh7 cells.These results suggest that CCNB1 may be an important gene to promote the proliferation of hepatoma cells.3.The expression levels of CCNB1 and CCNA2 were significantly correlated with the prognosis of patients with HCC.