医学分子生物学杂志2012，Vol.9Issue(2)：103-109,7.DOI:10.3870/j.issn.1672-8009.2012.02.006
靶向 IRE1a 干扰质粒构建及对 ERS 时细胞增殖及凋亡的影响
IRE1a-shRNA Plasmid Construction and Its Effect on Cell Prolifera-tion and Apoptosis
摘要
Abstract
Objective To construct the targeting IRE la mRNA interference plasmid pSUPER-IREla, study the influence of IRE la on cell proliferation and apoptosis of HeLa and HepG2 cells during ER Stress. Methods Two shRNA sequences specifically for IRE la were designed and constructed into recombinant plasmid pSUPER4REla, then transfected into human HeLa and HepG2 Cells. IREla gene expression levels in HeLa and HepG2 cells were detected with RT-PCR, Western blotting. Cell proliferation was examined by MTT and BrdU/DAPI immunofluorescence assays. Cell apoptosis was determined by flow cytometry. Results The recombinant plasmid pSUPER4REla downregulated IREla mRNA and protein levels in HeLa and HepG2 Cells. The cell proliferation rate and cell apoptosis rate were significantly different in experimental groups from control groups ( P < 0. 05 ) . Conclusion We successfully constructed the targeting IRE 1 a shRNA vector that inhibits mRNA and protein expression of IRE la in HeLa and HepG2 cells effectively. Besides, IREla-shR-NA promotes proliferation and inhibits apoptosis of HeLa and HepG2 cells during ER Stress. The IRE la shRNA vector can be used for further study of IRE la for its biological functions.关键词
IRE1a/shRNA/细胞增殖/细胞凋亡Key words
IRE1/ shRNA/ cell proliferation/ cell apoptosis分类
生物科学引用本文复制引用
姜蓉,刘艳娜,周菁华,刘平,郭风劲..靶向 IRE1a 干扰质粒构建及对 ERS 时细胞增殖及凋亡的影响[J].医学分子生物学杂志,2012,9(2):103-109,7.基金项目
国家自然科学基金 (No.81171697),重庆市自然科学基金 (No.2011jjA10047) （No.81171697）
