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RNA interference-based hTERT gene silence induces apoptosis of colorectal cancer SW480 cells
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Abstract:
Objective:To investigate the effect of RNA-mediated interference of hTERT (human telomerase reverse transcriptase) expression on the apoptosis of colorectal cancer cell line SW480. Methods：Small hairpin RNA (shRNA) targeting hTERT was synthesized and recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA (named hTERT-shRNA plasmid) containing hTERT-shRNA was constructed. SW480 cells were transfected with hTERT-shRNA plasmid by liposome method, and the expression of hTERT mRNA in SW480 cells was detected by RT-PCR analysis at different time points. The telomerase activity of SW480 cells was examined by TRAP-PCR-ELISA analysis. The ultrastructure of SW480 cells was examined by TEM (transparent electron microscope) 48 h after hTERT-shRNA transfection. Results: The inhibitory rate of hTERT mRNA expression in SW480 cells of hTERT-shRNA group was significantly higher than those of blank group, liposome group, and NC-shRNA group (75.0% vs 39.2%, 33.3%, 28.0%, P<0.05). Telomerase activity in SW480 cells of hTERT-shRNA group was significantly decreased compared with those of blank group, liposome group, and NC-shRNA group (2.242±0.285 vs 2.756±0.089, 2.693±0.225, 2.691±0.120, P<0.05). SW480 cells in hTERT-shRNA group showed smaller cell size, nuclear condensation, uneven aggregation of chromatin along the nuclear membrane, and increased vacuolization. Conclusion：RNA interference can effectively silence hTERT expression, reduce telomerase activity, and induce apoptosis of SW480 cells.
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Project Supported:
Project supported by the Natural Science Foundation of Guangxi (No. 2010GXNSFA013238), and the Key Science Foundation of Health Bureau of Guangxi (No. Z200971)
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