Effect of siRNA-induced CDK2 Expression Suppression on Expression of RB, CyclinE and E2F1 in Hepatic Carcinoma Cells

Objective To investigate the effects of cyclin-dependent kinase 2(CDK2) expression suppression induced by small interfering RNAs(siRNAs) on mRNA expression of cell cycle related genes RB,CyclinE and E2F1 in hepatic carcinoma cells SMMC7721.Methods The siRNA eukaryotic expression plasmids of CDK2 gene were constructed firstly and then were transfected into SMMC7721 cells with the Lipofectmine TM 2000 liposome.The transfected cells were divided into six groups: recombinant plasmid 190 group,recombinant plasmid 191 group,SMMC7721 group,CDK2-siRNA transfection group,negative control group,and blank vector group.The expression of CDK2 gene was detected with Western blot method.Real-time fluorescent quantitation polymerase chain reaction(PCR) method was utilized to detect the mRNA expression of RB,Cyclin E and E2F1 which were related to CDK2 gene,and then the effective siRNA sequence of CDK2 gene was screened.Results After the siRNA eukaryotic expression plasmids of CDK2 gene was transfected into SMMC7721 cells,mRNA expression of RB was up-regulated and the mRNA expression of CyclinE and E2F1 was down-regulated.Conclusion CDK2 gene expression suppression can up-regulate the mRNA expression of RB in SMMC7721,and down-regulate the mRNA expression of CyclinE and E2F1,indicating that the mRNA expression of RB,Cyclin E and E2F1 genes is correlated with CDK2 gene expression.