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腺病毒介导的血管平滑肌细胞和内皮细胞的体外基因转移。

Adenovirus mediated gene transfer of vascular smooth muscle cells and endothelial cells in vitro.

作者信息

Zhou H, Zeng G, Zhou A, Tang J, Huang Q, Chen G, Peng T, Hu B

机构信息

Cardiovascular Research Institute, Beijing Medical University.

出版信息

Chin Med J (Engl). 1995 Jul;108(7):493-6.

PMID:7555265
Abstract

Introducing foreign gene(s) into vascular smooth muscle cells (VSMCs) and endothelial cells (ECs) is the pre-requirement of gene therapy for cardiovascular diseases. We have explored the use of adenoviral vectors (Adv-CMV/LacZ) to transfer LacZ gene into cultured VSMCs and ECs. Our results demonstrated that adenoviral vectors transferred foreign gene into VSMCs and ECs high-efficiently with dose-dependent response pattern. The frequencies of transfection reached 100% at the viral titer of 10(9) pfu/ml. Comparing the sensitivities of VSMCs and ECs to adenoviral vectors, we found that ECs were more sensitive than VSMCs, of which the frequencies of transfection in ECs reached 80% while in VSMCs only 40% for 8 hrs after transfection. In addition, the transfection of ECs and VSMCs with adenoviral vectors was partly blocked by monoclonal antibodies to Fiber and Core protein of the adenoviral capsid, but not by monoclonal antibody to Hexon protein. It is suggested transfection of ECs and VSMCs with adenovirus vectors is mediated by Fiber or Core protein of adenoviral capsid proteins.

摘要

将外源基因导入血管平滑肌细胞(VSMCs)和内皮细胞(ECs)是心血管疾病基因治疗的前提条件。我们探索了使用腺病毒载体(Adv-CMV/LacZ)将LacZ基因导入培养的VSMCs和ECs。我们的结果表明,腺病毒载体能高效地将外源基因导入VSMCs和ECs,且呈剂量依赖性反应模式。在病毒滴度为10(9) pfu/ml时,转染频率达到100%。比较VSMCs和ECs对腺病毒载体的敏感性,我们发现ECs比VSMCs更敏感,转染8小时后,ECs中的转染频率达到80%,而VSMCs中仅为40%。此外,腺病毒载体对ECs和VSMCs的转染部分被针对腺病毒衣壳纤维蛋白和核心蛋白的单克隆抗体阻断,但未被针对六邻体蛋白的单克隆抗体阻断。这表明腺病毒载体对ECs和VSMCs的转染是由腺病毒衣壳蛋白的纤维蛋白或核心蛋白介导的。

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