嵌合性硫代磷酸修饰的反义c-myc寡脱氧核苷酸对动脉平滑肌细胞迁移的影响

目的：血管平滑肌细胞（ＳＭＣ）迁移在血管成形术后再狭窄的发生中起着关键作用。ｃ－ｍｙｃ是对多种刺激ＳＭＣ迁移的生长因子作出迅速早期反应的基因。推测采用与ｃ－ｍｙｃｍＲＮＡ互补的寡脱氧核苷酸（ＯＤＮ）的反义战略可达到抑制ＳＭＣ迁移的目的。方法：将嵌合性硫代磷酸修饰的反义ｃ－ｍｙｃＯＤＮ通过ｌｉｐｏｆｅｃｔｉｎ导入ＳＭＣ后，采用改良的Ｂｏｙｄｅｎｃｈａｍｂｅｒ方法检测ＳＭＣ的迁移率。结果：反义ｃ－ｍｙｃＯＤＮ作用的ＳＭＣ对１０％ＦＢＳ的化学趋化活性明显减弱，反义ｃ－ｍｙｃＯＤＮ显著抑制了ＳＭＣ迁移。而正义ｃ－ｍｙｃＯＤＮ和错配ｃ－ｍｙｃＯＤＮ处理的ＳＭＣ，与对照组相比，迁移率无差别。结论：（１）反义ｃ－ｍｙｃＯＤＮ对体外ＳＭＣ迁移的抑制具有序列特异性；（２）ｃ－ｍｙｃ基因表达在ＳＭＣ迁移的信号通路中起关键作用；（３）本研究为采用ｃ－ｍｙｃＯＤＮ的战略抑制ＳＭＣ迁移进而防止血管成形术后再狭窄的发生奠定了基础

The effect of chimeric antisense phosphorothioated c-myc oligodeoxynucleotide on cultured rat smooth muscle cell migration

AIM: Vascular smooth muscle cell (SMC) migration plays a pivotal role in restenosis following angioplasty. c -myc is an immediate early response gene induced by various growth factors, many of which have been found to stimulate SMC migration. It is reasoned that antisense oligodeoxynucleotide (ODN) complementary to c -myc mRNA might be potent inhibitors of SMC migration. METHOD: To confirm the hypothesis, chimeric antisense c -myc phosphorothioated ODN was introduced into cultured rat SMC by lipofectin. Migration activity was assayed by modified micro Boyden chamber method. RESULTS: The chemotactic activity of SMC treated with antisense c -myc ODN was inhibited significantly compared with that of other groups, whereas there was not a significant difference between control and mismatch or sense group. CONCLUSIONS: These results suggested:(1)Antisense c -myc ODN inhibits SMC migration in a sequence-specific manner; (2)c -myc product is involved in signalling transduction pathways mediating SMC migration in vitro ;(3) The c -myc antisense approach might find its application in preventing restenosis after angioplasty.