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siRNA interference of CHRNA5 gene expression inhibits nicotine-induced proliferation of human lung cancer cells
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Abstract:
Objective : To study the effect of siRNA targeting CHRNA5 (cholinergic receptor nicotinic alpha 5) gene expression on nicotine-induced proliferation of lung cancer A549 cells. Methods: CHRNA5-siRNA fragment was designed and synthesized, and then transfected into A549 cells by Lipofectamine assay. Expressions of CHRNA5 mRNA and protein in transfected-A549 cells were examined by FQ-PCR and Western blotting analysis, respectively. The effect of CHRNA5-siRNA on nicotine-induced proliferation of A549 cells was detected by MTT method. Results: CHRNA5-siRNA was successfully constructed and transfected into A549 cells. CHRNA5 mRNA expression in CHRNA5-siRNA-transfected A549 cells was significantly lower than that in control si-NC control siRNA transfected A549 cells (0.196±0.044 vs 0.944±0027, P<001), and CHRNA5 protein expression in CHRNA5-siRNA-transfected A549 cells was also lower than that in si-NC-transfected A549 cells (0.267±0.031 vs 0.745±0.035, P<0.01). When A549 cells were not treated with nicotine, CHRNA5-siRNA transfection inhibited the proliferation of A549 cells. But Nicotine increased the proliferation of A549 cells (P<0.01), and this positive effect was inhibited by CHRNA5-siRNA transfection (P<0.01). Conclusion: siRNA targeting CHRNA5 expression can inhibit nicotine-induced proliferation of A549 cells, which suggests that CHRNA5 may serve as a potential target for gene therapy of lung cancer.
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Project Supported:
Project supported by the Special Scientific Foundation for Postdoctoral in China (No.200902575), and the Start Foundation for Returned Chinese Scholars of Ji’nan(No.20080407)
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