中国病理生理杂志2012，Vol.28Issue(12)：2182-2186,5.DOI:10.3969/j.issn.1000-4718.2012.12.013
靶向COL1A1基因的 shRNA对人乳腺癌MDA-MB-231细胞增殖与凋亡的影响
Effects of shRNA targeting COL1A1 gene on proliferation and apoptosis of human breast cancer cell line MDA-MB-231
摘要
Abstract
AIM; To investigate the effects of shRNA -mediated collagen type I alpha 1 (COL1A1) gene silencing on the proliferation and apoptosis of human breast cancer cell line MDA - MB - 231. METHODS: The specific re-combinant vector pSilencer2.1 - U6 - COL1 Al was transiently transfected into human breast cancer cell line MDA - MB -231 with lipofectamine. RT -PCR and Western blotting were performed to detect the expression levels of COL1A1. MTT assay was employed to evaluate the effect of COL1A1 gene silencing on the cell proliferation. Flow cytometry was used to determine the apoptosis and cell cycle of transfected cells . The morphological characteristics of apoptosis were observed by Hoechst 33258 staining. RESULTS: Compared with mock group and scrambled group , the mRNA and protein levels of COL1A1 were reduced by pshRNA -COL1A1 transfection (P<0.05). The proliferation of MDA -MB-231 cells treated in shRNA - COL1 Al was significantly inhibited in a time - dependent way. The percentages of G0/G, phase cells and early apoptotic rate were significantly higher in pshRNA - CO LI Al group than those in mock and scrambled group ( P < 0.05 ). The changes of apoptotic morphology such as cell shrinkage and nuclear condensation were also observed by staining with Hoechst 33258 under fluorescence microscope. CONCLUSION: Transfection of eukaryotic expression vector pshRNA -COL1 Al effectively inhibits the proliferation , induces apoptosis and arrests MDA - MB - 231 cells in G0/G, phase.关键词
RNA干扰/I型胶原α1链/乳腺肿瘤/细胞凋亡Key words
RNA interference/Collagen type I, alpha 1 chain/Breast neoplasms/Apoptosis分类
医药卫生引用本文复制引用
余海浪,刘安玲,周珏宇,孟伟,郑文岭,马文丽..靶向COL1A1基因的 shRNA对人乳腺癌MDA-MB-231细胞增殖与凋亡的影响[J].中国病理生理杂志,2012,28(12):2182-2186,5.基金项目
广东省医学科研基金资助项目(No.B2012203) （No.B2012203）
