ADAM9特异性siRNA对肾透明细胞癌786-0细胞ADAM9基因表达及体外侵袭能力的影响

Effect of ADAM9 targeted siRNA on ADAM9 gene expression and invasion capacity of renal clear cell carcinoma 786-0 cells in vitro

目的:探讨去整合素金属蛋白酶9(ADAM9)特异性siRNA对人肾透明细胞癌(RCCC)786-0细胞中ADAM9基因表达及体外侵袭能力的影响。方法:设计合成ADAM9特异性siRNA。将786-0细胞分4组处理,分别为正常对照组(正常培养786-0细胞)、空脂质体转染组、阴性转染组(转染无义siRNA)和ADAM9 siRNA转染组。转染24、48、72 h后,采用RT-PCR法检测细胞ADAM9 mRNA的表达;转染48 h后,采用Western blot法检测细胞ADAM9蛋白的表达,并用Transwell法检测细胞侵袭能力。结果:转染24、48、72 h后,ADAM9 siRNA转染组786-0细胞ADAM9 mRNA的表达显著低于其他3组(F=47.945、180.456、60.978,P均<0.001);转染48 h后,ADAM9 siRNA转染组786-0细胞ADAM9蛋白表达显著降低(F=142.816,P<0.001),穿膜细胞数显著减少(F=45.389,P<0.001)。结论:ADAM9特异性siRNA能够有效沉默786-0细胞中ADAM9的表达并降低其体外侵袭能力。

Aim：To explore the effect of a disintegrin and metalloprotease 9 （ ADAM9 ） targeted siRNA on ADAM9 gene expression and invasion capacity of renal clear cell carcinoma 786-0 cells.Methods：ADAM9 specific siRNA was de-signed and synthesized .786-0 cells were divided into 4 group：normal cultured group , Lipofectamine TM 2000 transfection group, unrelated sequence siRNA （NC siRNA）transfection group and ADAM9 specific siRNA transfection group.RT-PCR was applied to detect ADAM9 mRNA in 786-0 cells after transfection for 24,48,and 72 h.ADAM9 protein was detected by Western blot after transfection for 48 h, and cell invasion was detected with Transwell .Results：The expression of ADAM9 mRNA in ADAM9 specific siRNA transfection group after transfection for 24,48, and 72 h were lower （ F =47.945,＆amp;nbsp;180.456,60.978,P＆lt;0.001）,and ADAM9 protein expression and invasion capacity were significantly lower after transfec-tion for 48 h（F=142.816,45.389,P＆lt;0.001）.Conclusion：ADAM9 specific siRNA can silence ADAM9 gene in 786-0 cells and inhibit its invasion .