Enhanced chemotherapy sensitivity of human colon cancer cells to 5-fluorouracil by siRNA recombinant expression vector targeting survivin gene.
- Author:
Ming CAI
1
;
Guo-bin WANG
;
Kai-xiong TAO
;
Chang-xue CAI
Author Information
- Publication Type:Journal Article
- MeSH: Antimetabolites, Antineoplastic; therapeutic use; Cell Line, Tumor; Cell Proliferation; Colonic Neoplasms; drug therapy; genetics; metabolism; Fluorouracil; therapeutic use; Genetic Vectors; genetics; metabolism; Humans; Inhibitor of Apoptosis Proteins; Microtubule-Associated Proteins; genetics; metabolism; RNA, Small Interfering; genetics; metabolism
- From: Chinese Medical Sciences Journal 2009;24(2):97-101
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo investigate the effects of small interfering RNA (siRNA) recombinant expression vector targeting survivin gene on chemotherapy sensitivity of human colon cancer cells to 5-fluorouracil.
METHODSsiRNA recombinant expression vector targeting survivin gene was constructed and transfected into human colon cancer cell lines LOVO. After 48 hours of transfection, cells were harvested for analysis of survivin mRNA and protein expressions using RT-PCR and Western blot. In addition, after human colon cancer cell lines were treated with Survivin siRNA and/or 5-fluorouracil, MTT assay and flow cytometry were used to analyze cell proliferation and apoptosis.
RESULTSRestriction endonuclease analysis confirmed that siRNA recombinant expression vector targeting survivin gene was successfully constructed. Inhibitory ratios of survivin mRNA and protein expressions by Survivin siRNA were 36.33% and 44.65%, respectively. Survivin siRNA combined with 5-fluorouracil significantly increased the cell proliferation inhibitory ratio and apoptosis ratio compared with 5-fluorouracil treating alone (P<0.05).
CONCLUSIONThe siRNA recombinant expression vector targeting survivin gene can inhibit the expression of survivin gene, and enhance chemotherapy sensitivity of human colon cancer cells to 5-fluorouracil.