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孙 旗 徐成斌
摘 要 目的：探讨逆转录病毒介导的N-ras 1基因对血管紧张素Ⅱ诱导的心肌细胞增殖和肥大的抑制作用，及其应用于心肌肥厚基因治疗的可能性。方法：将重组逆转录病毒反义N-ras 1基因体外转染乳鼠心肌细胞。应用RT-PCR及Western印迹法检测基因表达情况。应用细胞计数、［3H］-TdR掺入及细胞体积检测手段，观察N-ras 1基因对心肌细胞增殖及肥大的抑制作用。结果：逆转录病毒载体能将外源性基因导入心肌细胞并表达。反义N-ras 1基因导入可抑制心细胞增殖及肥大。结论：逆转录病毒介导N-ras 1基因有可能作为心肌肥厚的基因治疗手段。
主题词 基因；心肌；细胞， 培养的；血管紧张素Ⅱ；原癌基因
Inhibitory effect on antisense N-ras 1 gene on the proliferation and hypertrophy of cardiomyocytes induced by angiotension Ⅱ
SUN Qi, XU Cheng-Bin
Department of Cardiology, People s Hospital, Beijing Medical Univrersity, Beijing(100044)
Abstract AIM:To study the inhibitory effect of antisense N-ras 1 gene mediated by retrovirus vector on proliferation and hypertyophy of cardiomyocytes induced by Ang Ⅱ and the possibility by using antisense N-ras 1 gene for gene therapy of cardiac hypertrophy. METHODS：Antisense N-ras 1 gene recombinant retrovirus vector was transfected into cardiomyocytes in vitro. The expression of ras mRNA and p21 protein were detected by RT-PCR and Western blotting. The inhibitory role of antisense N-ras 1 gene for proliferation and hypertrophy of cardiomyocytes was observed by cell counting,［3 H］-TdR incorporation and cell volume respectively. RESULTS：Retrovirus vector tramsfered antisense N-ras 1 gene into cardiomyocytes. The expression of antisense N-ras 1 gene restricted the proliferation and hypertrophy of cardiomyocytes. CONCLUSION：Antisese N-ras 1 gene mediated by retrovirus can be used to treat cardiac hypertrophy.
MeSH Genes; Myocardium; Cells, cultured; Angiotensin Ⅱ; Prot-oncogenes
心肌肥厚是心肌及其间质细胞对生长因子的一种应答反映，是体内一系列基因异常表达的结果。血管紧张素Ⅱ(angiotension Ⅱ,Ang Ⅱ)是影响心肌肥厚的主要因子，具有促心肌细胞增殖和肥大的作用［1］。原癌基因ras表达产物p21蛋白是一种小分子量的G蛋白，可与GTP结合向细胞内传递增殖信息［2］。因而Ang Ⅱ的作用可能与p21蛋白有关。本研究选用逆转录病毒载体重组反义N-ras 1基因体外传染乳鼠心肌细胞，观察对心肌细胞增殖和肥大的作用，以探讨利用反义技术治疗心肌肥厚的可能性。
材料与方法
一、材料：
N-ras 1质粒由上海肿瘤研究所提供。基因片段长度0.9 kb，氨苄抗性。包装细胞PA317和NIH3T3由中科院病毒所提[1] [2] 下一页
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