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Effect of RNAi-mediated silencing of the human telomerase reverse transcriptase gene on colorectal cancer cell proliferation in vitro
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Abstract:
Objective: To construct an optimized human telomerase reverse transcriptase (hTERT) gene-specific RNAi and to evaluate its effect on human colon cancer cell proliferation in vitro. Method: Three hTERT-specific RNAi sequences and a negative control (NC) or scrambled sequence were cloned, respectively, into a pGPU6/GFP/Neo vector to generate pGPU6-GFP-hTERT-1, pGPU6-GFP-hTERT-2, pGPU6-GFP-hTERt-3 and pGPU6-GFP-NC. Human colon cancer SW480 cells were transfected with these vectors respectively. At 24, 48 and 72 h after transfection, hTERT mRNA abundance was assessed by RT-PCR and cell viability by MTT assay. Results: The 3 hTERT-specific RNAi vectors constructed were all effective to silence the hTERT gene; hTERT mRNA abundance in SW480 cells transfected with pGPU6-GFP-hTERT-3 was significantly lower than that in SW480 cells transfected with pGPU6-GFP-NC (0.347±0.028 vs 0513 ± 0.032,P<0.01). All the three hTERT sequence-specific RNAi vectors were effective to inhibit the proliferation of SW480 cells; cellular proliferation inhibition rate in SW480 cells of pGPU6-GFP-hTERT-3 group was significantly increased than that of blank contro, liposomal and NC group (\[50.08±0.43\]% vs \[4.11±0.39\]%, \[3.88±035\]% and \[3.38±0.35\]%; P<0.05). Conclusion: RNAi-mediated hTERT gene silencing results in colon cancer cell growth inhibition and may offer a novel therapy for colon cancer.
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Project Supported:
Project supported by the Shandong Wanjie Medical School Subject (No. X10ZK02)
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