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

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. Key words: MicroRNAs; Colonic neoplasms; Cell proliferation; Survivin