Journal of International Oncology››2015,Vol. 42››Issue (1): 22-26.doi:10.3760/cma.j.issn.1673-422X.2015.01.006

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Construction and inhibitory effect of microRNA expression vector targeting survivin on proliferation of human colorectal carcinoma HT-29 cells

Cui Yunfu, Hao Tao, Wang Ronghua, Li Baosong, Ma Chong, Fan Peng

  1. Department ofAnorectal Surgery,Affiliated Hospital of Binzhou Medical University, Binzhou 256603, China
  • Received:2014-08-18Revised:2014-10-08Online:2015-01-08Published:2015-01-07
  • Contact:Hao Tao E-mail:anmly521@163.com

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. MethodsmiRNA 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. ResultsThe 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). ConclusionSurvivin targeted silence can effectively decrease the expression of survivin mRNA and protein, induce colorectal carcinoma HT29 cell apoptosis and inhibit cell proliferation.

Key words:MicroRNAs,Colonic neoplasms,Cell proliferation,Survivin