E2F4对CDK2和CyclinA在肝癌细胞系中的转录调控特异性研究

目的研究E2F4对细胞周期素依赖性激酶2(CDK2)和细胞周期素A(CyclinA)在肝癌细胞系中的转录调控活性。方法扩增并克隆不同片段长度的CDK2和CyclinA启动子序列,E2F4表达质粒序列,转染肝癌细胞系(HepG2、QGY1007),双荧光报告系统检测荧光素酶表达活性,实时定量反转录-聚合酶链反应检测肝癌细胞系中CDK2和CyclinA mRNA的表达水平。结果双荧光报告系统、实时定量反转录-聚合酶链反应显示,在肝癌细胞系中转染E2F4表达质粒组CDK2和CyclinA启动子活性明显低于未转染E2F4表达质粒组,CDK2和CyclinA mRNA的表达水平降低。结论E2F4在肝癌细胞系中可以抑制CDK2和CyclinA的转录活性。

Evaluation of E2F4 Regulate CDK2 and CyclinA in Livet Cancer Cells

Objective To identify and evaluate the E2F4 regulation Cyclin-dependent protein kinase 2 (CDK2)and CyclinA in liver cancer cells.Methods Different sequences CDK2 and cyclinA gene promoters,E2F4 expression plasmids were amplified and constructed,were transfected into liver cancer cells(HepG2、QGY1007),the luciferase were detected to evaluate the E2F4 regulation activeties of CDK2 and cyclinA promoters,the real time-PCR were detected to evaluate the expression of CDK2 and cyclinA mRNA lever.Results The luciferase and real-time polymerase chain reaction assay the CDK2 and CyclinA promoters were transfected into E2F4 plasmids liver cancer cells were weaker activity than untransfected cells,the expression of CDK2 and CyclinA mRNA were decress than untransfected cells.Conclusion E2F4 represses activity of the CDK2 and CyclinA in liver cancer cells.