Construction and inhibitory effect of microRNA expression vector targeting survivin on proliferation of human colorectal carcinoma HT-29 cells
10.3760/cma.j.issn.1673-422X.2015.01.006
- VernacularTitle:靶向survivin基因微小RNA真核表达载体的构建及其对人结肠癌细胞增殖的影响
- Author:
Yunfu CUI
;
Tao HAO
;
Ronghua WANG
;
Baosong LI
;
Chong MA
;
Peng FAN
- Publication Type:Journal Article
- Keywords:
MicroRNAs;
Colonic neoplasms;
Cell proliferation;
Survivin
- From:
Journal of International Oncology
2015;42(1):22-26
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct microRNA (miRNA) expression vector targeting surviving,and to investigate its effect on transfected human colorectal carcinoma (HT-29) cell apoptosis and proliferation.Methods miRNA targeting survivin was synthesized and transfected HT-29 cells by lipofectin.HT-29 cells were cultured in the 6 orifices.The cultured cells were divided into control,liposome,negative control and positive control groups.Transient transfected cells were collected and the proliferation index and apoptosis rate of HT-29 cells were detected by flow cytometry.The expressions of survivin mRNA and protein were detected by RT-PCR and Western blot.Results The proliferation index and apoptosis rate of the positive control group were significantly higher compared with normal group,transfection group and mock-vehicle group (17.98％ ± 2.35％ vs 38.04％ ±2.11％ vs 36.73％ ±2.51％ vs 36.57％ ±3.05％; t =20.05,P＜0.01; t =18.75,P＜0.01; t=18.59,P＜0.01; 19.54％ ±1.74％ vs 3.13％ ±0.29％ vs 3.70％ ±0.44％ vs 3.61％ ± 0.50％ ; t =16.40,P ＜ 0.01 ; t =15.84,P ＜ 0.01 ; t =15.92,P ＜ 0.01).Survivin mRNA and protein expression levels were specifically suppressed in transfected HT-29 cells (t =0.68,P ＜0.01 ; t =0.58,P ＜ 0.01; t=0.61,P＜0.01;t=0.64,P＜0.01; t=0.62,P＜0.01;t=0.67,P＜0.01).Conclusion Survivin targeted silence can effectively decrease the expression of survivin mRNA and protein,induce colorectal carcinoma HT-29 cell apoptosis and inhibit cell proliferation.
