反义CD151基因转染对大鼠血管平滑肌细胞迁移的影响

目的观察pcDNA3.1真核表达载体介导的反义CD151基因转染对培养的大鼠动脉平滑肌细胞(VSMCs)迁移的影响。方法构建携带全长正义和反义CD151的真核表达载体pcDNA3.1-CD151和pcDNA3.1-anti-CD151重组质粒,转染体外培养的VSMCs,以RT-PCR和Western blot方法检测CD151的表达,用Boyden趋化小室方法观察细胞迁移。结果与载体对照组、脂质体对照组和空白对照组3组均值比较,转染48h后,反义CD151组mRNA表达降低58%,蛋白表达降低51%,正义CD151组的VSMCs CD151mRNA表达增加171%,蛋白表达增加133%;趋化迁移的细胞数,反义CD151组为37.9±6.3,正义CD151组为86.5±12.4;载体对照组、脂质体对照组和空白对照组分别为60.3±7.1、61.8±7.6和67.3±9.6。反义CD151组显著低于其余各组(P<0.01),正义CD151组显著高于其余各组(P<0.01)。结论pcDNA3.1真核表达载体介导的反义CD151转染,通过抑制CD151的表达,能够显著抑制大鼠VSMCs的迁移。

EFFECT OF ANTISENSE CD151 GENE TRANSFECTION TRANSFECTION ON THE MIGRATION OF VASCULAR SMOOTH MUSCLE CELLS IN RATS

Objective To explore the effect of the antisense sequence of CD151 gene transfection on the vitro migration of vascular smooth muscle cells(VSMCs) in rats.Methods Functional fragments of CD151 gene were amplified by RT-PCR and inserted into the vector pcDNA3.1 in the sense direction and antisense direction,respectively.Recombinant vectors pcDNA3.1-anti-CD151 and pcDNA3.1-CD151 were transfected into cultured VSMCs in vitro by means of lipofectamine. Their effects on mRNA and protein expression of CD151 were measured by RT-PCR and Western blot,and the effects on cell migration were evaluated by using Boyden chamber.Results In comparison with those of the controls,the CD151 expression was significantly decreased by 58% in mRNA and by 52% in protein,and the cell migration was significantly inhibited in VSMCs transfected with pcDNA 3.1-anti-CD151(P<0.01);but in VSMCs transfected with pcDNA 3.1-CD151,the CD151 expression was significantly increased by 171% in mRNA and by 133% in protein,and the cell migration was markedly enhanced as compared with those of the controls(P<0.01).Conclusion CD151 plays an important role in regulating cell migration in VSMCs.Antisense CD151 can attenuate the expression of CD151 and restrain the cell migration,which may provide a new gene target for anti-sense gene therapy in preventing and treauing restenosis after angioplasty.