腺病毒载体介导的胞嘧啶脱氨酶/单纯疱疹病毒-1-胸苷激酶双自杀基因对血管平滑肌细胞增殖的影响  

Effect of recombinant adenovirus expressing CD/TK double suicide genes on proliferation of vascular smooth muscle cells

作　　者：曹慧青[1] 孟宪敏[1] 赵勇[1] 赵秀文[1] 丁金凤[1] 

机构地区：[1]中国医学科学院中国协和医科大学阜外心血管病医院分子医学中心,北京100037

出　　处：《中华心血管病杂志》2002年第9期559-562,共4页Chinese Journal of Cardiology

基　　金：国家九五攻关课题 (96 90 6 0 2 0 7) ;国家自然科学基金课题 (3 9970 2 97)

摘　　要：目的　观察构建的含胞嘧啶脱氨酶 /单纯疱疹病毒 - 1-胸苷激酶 (CD TK)双自杀基因的重组腺病毒对血管平滑肌细胞 (VSMCs)增殖的影响。方法　将含CD TK单、双自杀基因的重组腺病毒感染培养至 3～ 5代的大鼠VSMCs。荧光显微镜观察报告基因绿色荧光蛋白的数量确定感染效率。用MTT法检测前体药物对各转基因组细胞增殖的抑制作用。结果　单、双自杀基因组腺病毒均可 10 0 %感染VSMCs。当感染复数 (M O I)为 10时 ,双自杀基因组靶细胞的存活率仅 10 % ,明显低于腺病毒 (Ad) TK(43% )和Ad CD(6 4 % )单基因组 (P <0 0 5 )。当M O I≤ 2 0时双自杀基因能够对细胞产生最大 (90 % )杀伤而CD、TK单基因组和单基因共转染组则不能 ,而且双基因组所需前体药物的浓度 (无环鸟苷 0 1μg ml,5 氟胞嘧啶 10 μg ml)低于单基因组 (无环鸟苷 0 5 μg ml,5 氟胞嘧啶 5 0 μg ml)。双基因组的TK或CD基因单独作用时对细胞的抑制作用接近或低于相应的单基因组。结论　双自杀基因共表达腺病毒载体对VSMCs的抑制作用、安全性及可调控性均优于单基因。Objective To study the effect of recombinant adenovirus expressing cytosine deaminase/ Herpes simplex virus 1 thymidine kinase (CD/TK) suicide gene on proliferation of vascular smooth muscle cells (VSMCs) Methods Rat VSMCs of 3 5 passages were infected by recombinant adenoviruses Ad TK, Ad CD, Ad TK CD and Ad lacZ The infection efficiency was measured by green fluorescent protein (GFP) detection The antiproliferation effect was determined by MTT assay Results (1)100% VSMCs could be infected with the recombinant adenovirus carrying either single or double gene The survival rate of VSMCs in Ad CD TK group under the M O I of 10 was reduced to 10%, lower than that of Ad TK(43%) and Ad CD(64%) groups( P <0 05) (2) When M O I≤20, the inhibition levels of VSMCs infected with CD/TK single suicide gene or co infected with both single genes reached their saturation, while the double gene group has most powerful inhibition effect (3) The concentration of prodrugs needed in double gene group (GCV 0 1μg/ml,5 Fc 10μg/ml)were lower than that in single gene groups(GCV 0 5 μg/ml,5 Fc 50μg /ml) when producing the highest killing effect Conclusion Recombinant adenovirus expressed double suicide gene had more powerful killing effect, was safer and easier to be regulated compared with the single gene

关 键 词：腺病毒载体介导 胞嘧啶脱氨酶 单纯疱疹病毒-1 胸苷激酶 自杀基因 血管平滑肌细胞 细胞增殖 

分 类 号：R456[医药卫生—治疗学]