Effect of microRNA-mediated p65 gene silencing on the proliferation and apoptosis of human breast cancer MDA-MB-231 cells.
- Author:
Ling WANG
1
;
Bao-en SHAN
;
Mei-xiang SANG
;
Yi-shui LIAN
;
Bin WANG
;
Chun-yan DING
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; genetics; Breast Neoplasms; genetics; pathology; Cell Line, Tumor; Cell Proliferation; Female; Gene Knockdown Techniques; Gene Silencing; Humans; MicroRNAs; genetics; RNA, Messenger; genetics; metabolism; Transcription Factor RelA; genetics; metabolism; Transfection
- From: Journal of Southern Medical University 2011;31(10):1742-1747
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the effect of microRNA (miRNA)-mediated p65 gene knockdown on the proliferation and apoptosis of human breast cancer MDA-MB-231 cells.
METHODSp65-targeted miRNA plasmid was constructed and transfected into MDA-MB-231 cells via lipofectamine(TM)2000. The expression of p65 gene in the transfected cells at the mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The cell proliferation and apoptosis were measured by MTT assay and flow cytometry (FCM), respectively. The expressions of apoptosis-related proteins were detected by Western blotting in the transfected cells.
RESULTSCompared with the negative control group, the expressions of p65 mRNA and protein in p65miRNA-trasnfected cells were significantly down-regulated (P<0.05). MTT assay showed significantly lowered viability of MDA-MB-231 cells after the transfection (P<0.05). FCM showed an increased cell apoptosis rate in p65miRNA group compared with that in the negative control group (P<0.05). Caspase-3 and PARP were both cleaved into their active forms and the expression of these active forms was increased in p65miRNA group.
CONCLUSIONThe miRNA targeting p65 gene can inhibit the proliferation and induce apoptosis of breast cancer cells, and p65 gene might become a new target in gene therapy for human breast cancer.
