地尔硫卓对增殖血管平滑肌细胞的原癌基因表达的影响

Effect of Diltiazem on the expression of proto-oncogene in proliferated vascular smooth muscle cells

目的观察地尔硫卓(Dil)对增殖血管平滑肌细胞(VSMC)的原癌基因c-myc、c-fos、c-jun和ras mRNA表达的影响。方法将组织贴块法培养的大鼠胸主动脉VSMC随机分为5组,即空白组、模型组和Dil1、2、3组(浓度分别为10-5、10-6、10-7mol/L),应用MTT检测增殖能力,用流式细胞术检测VSMC的增殖指数,用rt-PCR检测c-myc、c-fos、c-jun和ras mRNA的表达。结果与模型组比较各浓度Dil都能抑制VSMC增殖,PI值均显著下降(各组PI值分别为21.53±1.72、28.63±0.96、15.95±0.37、19.28±0.94、20.33±0.67;P<0.05);Dil1、2组c-myc、c-fos、c-jun mRNA的表达显著减少(模型组和Dil1、2组的c-myc/GAPDH值分别为2.454±0.03、1.509±0.05、1.660±0.04,c-fos/GAPDH值分别为0.0046±0.0004、0.0023±0.0003、0.0038±0.0005,c-jun/GAPDH值分别为1.950±0.03、1.077±0.03、1.725±0.03;P<0.05),rasmRNA的表达显著增加(模型组和Dil1、2组的ras/GAPDH值分别为1.941±0.03、3.811±0.02、2.501±0.02;P<0.05)。结论Dil抑制VSMC增殖机制可能与下调原癌基因c-myc、c-fos、c-jun的表达和上调ras的表达有关。

Objective To investigate the effect of Diltiazem （Dil） on the expression of the proto-oncogene c-myc,c-fos,c-jun and ras mRNA on VSMC. Methods Initially, VSMC were cultured by tissue-sticking method. Then they were randomly divided into 5 groups： control group, model group, Dil groups （Dil divided into 10-2,10-6 and 10^-7 mol/L）. Finally, the cell proliferation ability was measured by MTr assay, the cell generation cycle and prolifation index were measured by Flow cytometery, and the expressions of c-myc ,c-fos,c-jun and ras mRNA were examined by rt-PCR. Results Dil inhibited the proliferation of VSMC slguificantly（P〈0.05）, down- regulated the proliferation index significantly （P〈0.05） in all the Dil groups, in group 1 and 2,Dil decreased the expression of c-myc, c-los and c-jun mRNA significantly （P〈0.05） and increased ras mRNA significantly （P〈 0.05 ）. Conclusion Dil inhibited the proliferation of VSMC, and the mechanism concerned with down- regulating the expression of c-myc,c-fos and c-jun and up-regulating the expression of ras.