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Effects of siRNA silencing livin expression on proliferation, apoptosis and invasion of human colon cancer cell line HT-29
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Abstract:
Objective：To explore the effects of small interference RNA (siRNA) targeting livin on the proliferation, apoptosis and invasion of human colon cancer cell line HT-29. Methods: Chemically synthetic double-strand siRNA targeting livin (livin-siRNA) was transfected into HT-29 cells, and then RT-PCR and Western blotting were used to detect the expression of livin mRNA and protein in HT-29 cells. MTT assay was performed to analyze the proliferation of HT-29 cells. The cell apoptosis and cell cycle distribution were analyzed by flow cytometry. The invasion assay and caspase-3 detective kit were used to detect the change of invasion and caspase-3 activity in HT-29 cells. Results: Forty-eight hours after transfection, there was a significant decrease in the expressions of both livin mRNA (0.073±0007 vs 0.395±0082, 0.423±0.025, 0.418±0.032, P<0.05) and livin protein (0.106±0.003 vs 0.456±0065, 0.473±0078、0.491±0.045, P<0.05) in the livin-siRNA group, compared with the blank and negative control and liposome groups. Ninety-six hours after transfection, the growth of HT-29 cells in the livin-siRNA group was significantly lower than that in the control and liposome groups (0.564±0.102 vs 0.833±0.127, 0.860±0.153，P<005), and the rate of apoptosis was obviously increased (\[16.5±2.8\] % vs \[2.4 ±0.5\]%, \[3.7±1.0\] %, P<005). The invasion assay demonstrated that the number of the migration cells was lower in the livin-siRNA group than in the control and liposome groups (31.3±4.5 vs 101.3±8.6, 97.4±7.8, P<0.05). The activity of caspase-3 in the livin-siRNA group was decreased compared with that in the control group (0.160 ±0.023 vs 0.347± 0.058, P<0.05). Conclusion: The siRNA silencing livin expression in HT-29 cells can suppress the proliferation, induce the apoptosis and inhibit the invasion of HT-29 cells.
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Project supported by the National Natural Science Foundation of China (No. 30960440)
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