Screening Of Differentially Expressed Genes In Hepatocellular Carcinoma HepG2 Cells After Silencing CDC25A Gene

Objective: To construct human hepatocellular carcinoma HepG2 cell model that stably silences the cell division cycle 25A(CDC25A)gene.Methods: To infect Human hepatoma HepG2 cells through constructing a lentivirus RNA interference(RNAi)vector that can silence the expression of CDC25 A gene.Then using real-time PCR and Western blot to verify the silencing efficiency of CDC25 A gene.Results: Human hepatocellular carcinoma HepG2 cells stably silenced CDC25 A gene was constructed and the expression levels of CDC25 A mRNA and protein was down-regulated.The silencing efficiency of CDC25 A was about50%.Conclusion: The use of lentiviral vector and RNAi technology can be successfully construct the human hepatoma HepG2 cell line stably silenced CDC25 A gene.Objective:To study the difference in gene expression of Hep G2 cells after silencing CDC25 A gene by using gene chip technology,and to search for related genes that may regulate upstream and downstream genes of CDC25 A.Methods:Using human whole genome microarray screen the different expressed genes in Hep G2 cells after silencing CDC25 A,and analysis it by Ingenuity Pathway Analysis.Then using real-time quantitative PCR to verify the expression of different expressed gene related to cell cycle in the result of gene chip.Results: There were 1453 differentially expressed genes in human hepatoma Hep G2 cells after gene silencing CDC25 A screened by Affymetrix human gene expression profiling microarray,including 789 up-regulated genes and 664 down-regulated genes.According to IPA,using classical signaling pathway to enrich the significantly different expressed genes,mainly involving IL-6 signaling pathway,cell cycle pathway,P53 signaling pathway,PI3K/AKT signaling pathway,Wnt/ ?-catenin signaling,c AMP mediated Signaling pathways,Toll-like receptor signaling pathways,etc.The result indicates that IL-6 signaling pathway was significantly inhibited and the cell cycle pathway was also suppressed.In addition,disease and function analysis showed that Progression of tumor was significantly inhibited.It confirmed that the cell cycle related genes of CCND1,CDKN1 A,CDC42,IL6,MAP4K4 and CXCL8 genes were down-regulated by q RT-PCR in Hep G2 cells with silencing CDC25 A gene,which was consistent with the gene chip results.Conclusions:The human gene expression profile microarray can be used to screen the different expressed genes in human hepatocellular carcinoma cells after silencing CDC25 A gene,which provided clues for exploring the effect of CDC25 A gene on hepatoma cell growth.