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Effects of small interference RNA-mediated TLR4 silencing on the proliferation and invasion of human lung adenocarcinoma A549 cells
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Abstract:
Objective:To investigate the effects of small interference RNA (siRNA)-mediated TLR4 silencing on proliferation and invasion of human lung adenocarcinoma A549 cells. Methods: Three TLR4-specific siRNAs (siRNA722, siRNA1673 and siRNA2560) and one negative control siRNA were designed and chemically synthesized. One siRNA that could most effectively silence the TLR4 gene in A549 cells was chosen from the three test siRNAs based on their capacity of lowering TLR4 mRNA and protein levels revealed by real-time PCR and Western blotting analysis respectively. A549 cells were transfected with this most effective siRNA and the control siRNA respectively. Proliferation and invasion capacity of the transfectants were assessed by CCK-8 assay and Transwell assay, respectively. Results: TLR4-siRNA1673 was the most effective siRNA which could significantly inhibit the expression of TLR4 at both mRNA (0.45±0.03 vs 0.83±0.02, P <0.01) and protein (0.23±0.06 vs 0.98±0.09) levels at 48 h after transfection into A549 cells as compared with negative control siRNA. The proliferation of the TLR4-siRNA1673-transfected A549 cells was significantly inhibited compared with the NS-siRNA-transfected A549 cells ( P <0.05). Similarly, the invasion capacity was significantly lower in the TLR4-siRNA1673-transfected A549 cells than in the NS-siRNA-transfected cells (23.60±2.88 vs 59.80±5.54, P <0.01). Conclusion: Sequence-specific siRNA may effectively silence TLR4 expression and inhibit the proliferation and invasion ability of human lung carcinoma cells in vitro . This preliminary observation suggests that TLR4 may serve as a putative therapeutic target for lung cancer.
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Project Supported:
Project supported by the National Natural Science Foundation of China（No.81202789），the Scientific Research Project for High Education from Education Bureau of Liaoning Province（No.2009A500）, and the Doctoral Foundation of Liaoning Province（No.20111134）
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