目的：探讨Toll样受体4（TLR4）对脂多糖（LPS）促进人肺癌SPCA1细胞增殖的影响。方法：采用LPS（10 μg/ml）刺激细胞，模拟慢性炎症的细胞微环境。将靶向TLR4基因的小干扰RNA（TLR4-siRNA）或阴性对照（NC-siRNA）通过脂质体介导转染人肺癌SPCA1细胞，24 h后加入10 μg/ml LPS。按转染siRNA和加入LPS情况，实验分为不做任何处理的Control组、NC+10LPS组以及TLR4-siRNA+10LPS组。采用Real-time PCR和流式细胞术检测SPCA1细胞中 TLR4 mRNA及蛋白的表达情况；采用CCK-8法和平板克隆形成实验检测细胞增殖能力，流式细胞术检测分析细胞周期分布情况。结果：与NC+10LPS组相比较，TLR4-siRNA+10LPS组细胞中TLR4mRNA及蛋白的表达水平显著下降（P<0.01）；与Control组和NC+10LPS组相比较， TLR4-siRNA+10LPS组SPCA1细胞的增殖明显减缓（P<0.01），TLR4-siRNA+10LPS组细胞克隆形成能力明显降低\[（4.50±1.89）vs (13.33±1.81）、（15.75±1.25）个，P<0.01\]；TLR4-siRNA+10LPS组细胞周期阻滞于G0/G1期\[（61.55±0.55）% vs (53.59±1.59) %、（51.72±0.77）%，P<0.01\]。结论： TLR4-siRNA能有效沉默SPCA1细胞中TLR4的表达，能够阻滞细胞的生长，抑制细胞的增殖。

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<001), 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 G0/G1 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.

国家自然科学基金资助项目（No.81202789）；辽宁省教育厅高等学校科研项目资助（No.2009A500）；辽宁省博士启动基金项目资助（No.20111134）。