siRNA干扰对乳腺癌(MCF-7)ATX表达和侵袭力的影响

Effect of silencing ATX by RNA interference on expression and invasive potential of human breast cancer cell lines

目的:研究针对自分泌运动因子(Autotaxin,ATX)的小干扰RNA(Small interfering RNA,siRNA)对人乳腺癌细胞ATX基因表达的影响,探讨RNA干扰技术治疗乳腺癌的前景。方法:参考人黑色素瘤ENPP2基因序列,设计合成ATX-siRNA及随机阴性对照。在阳离子脂质体的介导下转染人乳腺癌细胞MCF-7,转染后48h收集细胞,用半定量RT-PCR和western blot检测转染后ATX mRNA和ATX蛋白的表达,体外实验测定肿瘤细胞穿透matrigel的能力。结果:体外合成的siRNA在转染乳腺癌细胞48h后ATX mRNA和蛋白表达降低,转染48h后测定乳腺癌MCF-7细胞的侵袭力明显降低(P<0.01)。结论:ATX-siRNA能有效抑制人乳腺癌细胞株ATX基因和蛋白的表达,降低人乳腺癌细胞的侵袭力。

Objective：To study the effect of RNA interference （RNAi） on ATX-mRNA expression and the invasive petential of human breast cancer cell lines. Methods： Chemically synthesizeddouble stranded RNA（dsRNA ） targeting ATX was transfected into human breast cancer cell MCF-7 using SiPORT Lipid. The transfection efficiency was observed under fluorescence confocal microscope. Expression of ATX mRNA and protein was detected by reverse transcription polymerase chain reaction （RT-PCR） and Western blot. Cell penetrate matrigel capacity were determined by in vitro experiment. Results：ATX-siRNA effectively inhibited ATX- mRNA and protein expression （P〈0.05）, and degraded invasion capability of MCF-7 cells（P〈0.01 ）. Conclusion：ATX-siRNA inhibits ATX-mRNA and protein expression, and degrades invasion of human breast cancer cell.