Effect of siRNA to inhibit chemokine receptor CXCR4 expression on invasion capability of human lung cancer cells in vitro / 肿瘤

ABSTRACT

Objective: To study the inhibitory effect of chemokine receptor CXCR4-targeted small interference RNA (siRNA) on invasion capability of human lung cancer cell line (A549) in vitro. Methods: siRNA for specifically targeting chemokine receptor CXCR4 was designed using T7 RNA polymerase. A549 cell was cultured in a standard condition and was transfected with siRNA via lipofectamin AMINE 2000 mediation. The mRNA level of CXCR4 was detected by RT-PCR at 72 h after transfection. The expression of CXCR4 protein was detected by Western blotting. Invasion capability of A549 cells in vitro was evaluated by using Boyden chamber model. The cell cycle distribution was measured by flow cytometry and the proliferation capability was determined by MTT assay. Results: At 72 h after transfection of CXCR4 siRNA, the expression of CXCR4 mRNA and protein was down-regulated significantly. The invasion and proliferation capabilities of A549 cells decreased substantially. But CXCR4 siRNA transfection had no significant effect on the distribution of cell cycle. Conclusion: CXCR4 siRNA effectively down-regulates the expression of CXCR4 gene and decreased invasion capability of A549 cells in vitro.