现代检验医学杂志2011，Vol.26Issue(5)：6-10,5.DOI:10.3969/j.issn.1671-7414.2011.05.003
Cyclin D1-siRNA真核质粒的构建及对HepG2肝癌细胞增殖的影响
Construction of Cyclin D1 siRNA Vector and Effect on Proliferation of Human HepG2 Cancer Cells
王一 1连小云 1张玎 1王晖 1王岐山1
作者信息
- 1. 陕西省人民医院血液科,西安,710068
- 折叠
摘要
Abstract
Objective To explore the effect of cyclinDl gene blocking by siRNA on proliferation and cell cycle of HepG2 liver caicenoma cells. Methods Four pairs of DNA templates coding siRNA,synthesized against cyclinDl and cloned into the vector PGE-l?were identified by restriction ndonuclease digestion analysis,PCR and DNA sequencing cells were then transfected with these four PGE-1-siRNAs and the negative control. After G418 selection,RT-PCR and Western blot were used to detect the expression of cyclinDl gene. Cell growth curve were drived by MTT assays. Results Restriction en-donuclease digestion analysis and DNA sequencing results all showed that the target segments were cloned PGE-1 vector respectively after the postive-siRNA was chosen to transfected into HepG2 cells,the expression of cyclinDlmRNA and protein was marketly decreased. The results showed that after interfering,the HepG2 cells cell growth were signifencantly inhibited. Conclusion The cyclinDl-specific siRNA mediated by PGE-1 could effectively knockdown the expression of gene and inhibits the proliferation potential ability of HepG2 cells.关键词
细胞周期素D1/RNA干扰/人肝癌HepG2细胞/细胞周期/增殖Key words
RNA infection/human liver cancer cell HepG2/cyclinDl/cell cycle/cell proliferation分类
医药卫生引用本文复制引用
王一,连小云,张玎,王晖,王岐山..Cyclin D1-siRNA真核质粒的构建及对HepG2肝癌细胞增殖的影响[J].现代检验医学杂志,2011,26(5):6-10,5.
