Construction of COL1A1 short hairpin RNA vector and its effect on cell proliferation and migration of gastric cancer cells.
- Author:
Ai-qing LI
1
;
Jian-min SI
;
Yan SHANG
;
Li-hong GAN
;
Lei GUO
;
Tian-hua ZHOU
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line, Tumor; Cell Proliferation; Collagen Type I; genetics; metabolism; Genetic Vectors; Humans; Plasmids; genetics; RNA, Messenger; genetics; RNA, Small Interfering; genetics; Stomach Neoplasms; pathology; Transfection; Transformation, Bacterial
- From: Journal of Zhejiang University. Medical sciences 2010;39(3):257-263
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo construct COL1A1-targeted short hairpin RNA (shRNA) vector with pSilencer 4.1-CMV neo siRNA expression vector and to evaluate its effect on proliferation and migration of gastric cancer BGC-823 cells in vitro.
METHODSThree COL1A1-shRNA plasmids (COL1A1-shRNA-1, COL1A1-shRNA-2, COL1A1-shRNA-3), targeting different sites of COL1A1 gene, were constructed using pSilencer 4.1-CMV neo siRNA expression vector and transfected into gastric cancer BGC-823 cells. Real time quantitative RT-PCR and Western blot were performed to detect expression levels of COL1A1. MTT and Transwell migration assays were employed to evaluate the effects of COL1A1 gene silence on cell proliferation and migration.
RESULTThree recombinant plasmids targeting COL1A1 were constructed successfully. The expressions of COL1A1 in BGC-823 cells, including mRNA and protein levels, were significantly inhibited by the COL1A1-shRNA transfectants, which resulted in a clear reduction of cell proliferation and migration capacity.
CONCLUSIONThe COL1A1-shRNA can effectively knock down gene expression and inhibit proliferation and migration of gastric cancer BGC-823 cells.
