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Effect of Toll-like receptor 4 gene silencing by small interfering RNA on lipopolysaccharide-induced human lung carcinoma SPCA1 cell proliferation
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Abstract:
Objective: To determine the effect of Toll-like receptor 4 (TLR4) on lipopolysaccharide (LPS)-induced human lung carcinoma SPCA1 cell proliferation. Methods: SPCA1 cells were treated with LPS (10 μg/ml) to mimic a chronic inflammation microenvironment. TLR4 specific small interfering RNA (TLR4-siRNA) and a negative control interfering RNA (NC-siRNA) were transfected into SPCA1 cells by Lipofectamine. At 24 h after transfection, transfectants were treated with 10 μg/ml LPS. At 48 h after LPS treatment, TLR4 mRNA and protein levels were determined by Real-time PCR and FCM respectively, cell proliferation was assessed by CCK-8 assay and colony formation assay, and the cell cycle distribution was examined by FCM. Results: TLR4 mRNA and protein levels were significantly decreased (P<001), proliferation was significantly suppressed (P<0.01), and colony formation ability was significantly reduced (P<0.01) in SPCA1 cells transfected with TLR4-siRNA as compared with cells transfected with NC-siRNA. The proportion of cells arrested at the G0/G1 phase was significantly higher (P<0.01) in SPCA1 cells transfected with TLR4-siRNA (61.55±0.55)% than in NC-siRNA-transfected (53.59±1.59)% and control (51.72±0.77)%. Conclusion: TLR4-siRNA can effectively silence TLR4 expression in SPCA1 cells, block cell growth and inhibit cell proliferation.
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Project Supported:
Project supported by the National Natural Science Foundation of China（No.81202789）, the Scientific Research Project from the Bureau of Education of Liaoning Province（No.2009A500）, and the Doctoral Foundation of Liaoning Province（No.20111134）
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