小干扰RNA对骨肉瘤细胞株CXCR4基因表达和侵袭力的影响

Effects of small interfering RNA on CXCR4 expression and metastatic potention in osteosarcoma cell subline

目的:研究小干扰RNA(siRNA)对骨肉瘤细胞株中趋化因子受体CXCR4基因表达和侵袭力的影响。方法:采用单细胞克隆技术,从人骨肉瘤细胞系MG-63中分离培养出高表达CXCR4的骨肉瘤细胞株,设计合成针对CXCR4基因的siRNA,脂质体法转染骨肉瘤细胞株后用RT-PCR检测CXCR4基因表达水平的变化,荧光检测转染效率,Transwell趋化侵袭实验观察转染后细胞株侵袭能力的变化。结果:转染效率约为43.82%,CXCR4编码基因序列特异性siRNA能有效下调CXCR4基因表达水平,与空白组和阴性对照组比较,其差异有统计学意义(P<0.05),细胞株穿过侵袭膜的细胞数也较转染前减少(P<0.05)。结论:小分子干扰RNA技术能有效抑制骨肉瘤细胞株CXCR4基因的表达和体外侵袭能力。

Objective：To study the effects of small interfering RNA （siRNA） on the expression of CXCR4 and metastatic potention in osteosarcoma cell subline. Methods： We tried to obtain osteosarcoma cell sublines by the unicell cloning technique from human osteosarcoma cell line MG - 63, choose the cell subline of high expression of CXCR4 mRNA by RT - PCR, designed and synthesized one CXCR4 target sequence - specific small interfering RNA （siRNA） ,transfected siRNA into osteosarcoma cell subline by using Lipofectamine^TM 2000, observed the transfection efficiency and the change of expression of CXCR4 mRNA, detected the change of metastatic potention by Transwell vitro invasion assay. Results ： The expression of CXCR4 mRNA and the metastatic potention of osteosarcoma cell subline was effectively inhibited by CXCR4 - siRNA（ P 〈 0.05 ） ,the transfection efficiency was about 43.82%. Conclusion ： The CXCR4 - siRNA could successfully inhibit the expression of target gene and the vitro metastatic potention of osteosarcoma cells, and it can be potentially useful in the next research of pulmonary metastasis of osteosarcoma.