基因疗法__(期刊论文)

cmyc基因mRNA核酶对血管平滑肌细胞增殖的抑制作用

http://www.100md.com 《第四军医大学学报》 2003年第6期

基因疗法,,基因，myc；RNA，催化；肌，平滑，血管,细胞学；基因疗法,0引言,1材料和方法,2结果,3讨论,【参考文献】

     (第四军医大学：西京医院心血管内科，陕西西安 710033，唐都医院传染科，陕西西安 710038)

    Inhibition of vascular smooth muscle cell proliferation by specific cmyc mRNA cleaving hammerhead ribozyme

    LUAN RongHua, JIA GuoLiang, LI Wei, JIA ZhanSheng, LIAN JianQi

    Department of Cardiovasology, Xijing Hospital, Fourth Military Medical University, Xian 710033, China, Department of Infectious Diseases, Tangdu Hospital, Fourth Military Medical University, Xian 710038, China

    【Abstract】 AIM： To investigate the effects of hammerhead ribozyme that specifically cleaves cmyc mRNA on the proliferation of vascular smooth muscle cells (VSMCs). METHODS： Based on the computer analysis of the secondary structure of cmyc mRNA, nt 2029 in rat cmyc oncogene was selected as the cleaving site for hammerhead ribozyme and the ribozyme was designed. With an automatic DNA synthesizer, two complementary DNA strands of the ribozyme were synthesized. The ribozyme gene was cloned into pGEM3Zf (+) vector and subcloned into eukaryotic expression pcDNA3 vector. The recombinant pcDNARz was transfected into the cultured rat VSMCs by lipofectAMINE mediated DNA transfection protocol and individual cell clones were selected by G418. RESULTS： The sequence of ribozyme gene inserted in pGEM3Zf (+) vector was proved to be perfectly correct. In vascular smooth muscle cells transfected with recombinant pcDNARz, flow cytometry analysis showed that the S phase and G2/M fractions decreased significantly and VSMCs proliferation stagnated in the G0/G1 phase. CONCLUSION： The results suggest that hammerhead ribozyme that specifically cleaves cmyc mRNA can significantly inhibit the proliferation of VSMCs. ......

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