小干扰RNA沉默血管内皮生长因子对ACHN肾癌细胞黏附、侵袭及迁移的影响

Effects of small interference RNAs targeting VEGF on adhesion, migration and invasion of human renal cell cancer cell line ACHN

目的观察小干扰RNA（siRNA）沉默血管内皮生长因子（VEGF）对ACHN肾细胞癌细胞生物学行为的影响。方法化学合成针对VEGF的小干扰RNA，实验分4组，转染后收集细胞，通过蛋白印迹方法检测VEGF的表达，噻唑蓝（MTY）比色法、细胞黏附实验、划痕实验及Transwell法测定细胞的增殖、黏附、迁移及侵袭能力。结果siRNA1～2组VEGF表达水平明显低于空白对照组及阴性对照组；在24、48、72h，siRNA1—2的增殖抑制率均高于空白对照组及阴性对照组（P〈0．05），其中以siRNA2组最为显著；与空白对照组比较，siRNA1—2组的细胞黏附数量明显下降[（81．5±3．1）％比（40．54-2．6）％、P〈0．05，（81．5±3．1）％比（22．5±2．4）％、P〈0．05]，其中以siRNA2组最为明显；siRNA1～2组的细胞迁移数量明显下降（162±9比81±5，P〈0．05；162-1-9比38±4，P〈0．05）；siRNA1—2组细胞侵袭能力明显下降（P〈0．05），且siRNA2组的细胞侵袭能力明显低于siRNA1组（P〈0．05）。结论VEGF基因在肾细胞癌细胞黏附、迁移和侵袭中发挥着重要作用；以靶向VEGF的siRNA转染肾细胞癌细胞，可抑制肾细胞癌细胞黏附、迁移和侵袭能力。

Objective To study the effect of specific small interfering RNA （siRNA） silencing vascular endothelial growth factor （VEGF） on adhesion, migration and invasion of human renal cell cancer cell line ACHN. Methods The chemically synthetic siRNA targeting VEGF was transfected into ACHN cells, and four experimental groups were set up. The expression of VEGF was detected by using Western blotting, the inhibition of proliferation was studied by methyl thiazolyl tetrazolium （MTT） assay, the abilities of cell mobility and adhesion were detected by wound healing assay and adhesion test, and the abilities of cell invasion were evaluated by using Transwell chambers. Results After siRNA interference, the cells in siRNA1-2 groups transected with a specific VEGF-siRNA showed suppression of VEGF protein expression compared with that in the blank control group and negative control group, especially in siRNA2 group; the abilities of adhesion in siRNA1-2 groups, as compared with the blank control group, were decreased [（81.5±3.1%） vs. （40.5±2.6）%,P〈0.05; （81.5±3.1）% vs. （22.5±2.4）%,P〈0.05], especially in siRNA2 group; the abilities of migration in siRNA1-2 groups, as compared with the blank controi group, was decreased （162 ±9 vs. 81 ±5,P〈0.05; 162 ±9 vs. 38 ±4,P〈0.05）, especially in siRNA2 group; the abilities of invasion in siRNA1-2 groups were suppressed （P 〈 0.05 ） as compared with the blank control group, especially in siRNA2 group （ P 〈 0. 05 ）. Conclusion VEGF gene might play an important role in adhesion, migration and invasion of human renal ceil carcinoma cells, siRNA targeted VEGF could effectively inhibit adhesion, migration and invasion of human renal cell carcinoma cells.