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论文摘要

端粒酶催化亚单位转染血管内皮细胞的实验研究

Studies on the transfection of Umbilical Endothelia with Catalytic Subunit of Telemerase

作者:代晓明,李龙江,温玉明,王昌美,刘华,刘坤,李胜富

Author:DAI Xiao-nung1,LI Long- Jiang,WEN Yu-ming'.WANG Chang-mei1,LIU Hun1,LIU Kun1,LI Sheng fu2

收稿日期:2004-10-25          年卷(期)页码:2004,22(05):373-

期刊名称:华西口腔医学杂志

Journal Name:West China Journal of Stomatology

关键字:血管内皮细胞,端粒酶催化亚单位,转染,

Key words:vascular endothelium,catalytic subunit of telemerase,

基金项目:

中文摘要

目的探讨端粒酶催化亚单位转染血管内皮细胞后,血管内皮细胞增生活性的变化。方法应用胶原酶消化法培养脐静脉血管内皮细胞(UE),ABC法检测内皮细胞CD34表达。脂质体介导内皮细胞转染,MTT法测定细胞代谢活性并测定细胞生长曲线。基于逆转录聚合酶链反应的端粒重复末端扩增分析法检测端粒酶活性表达。结果培养的细胞贴壁后为单层呈铺路石状排列。细胞爬片CD34染色呈阳性,pBABE-HYGRO-hERT转染内皮细胞测定端粒酶活性的吸光度为0.889. NIT],实验表明:在各个时间点转染阳性细胞(HC)的吸光值都高于脐静脉血管内皮细胞(UE)(P

英文摘要

Objective To investigate the variety of proliferating ability of umbilical endothelia《UE) transfected场plasmid pBABE-HYGR-hTERT. Methods UE~identified from two aspects: morphology and CD34 labeling technique. The plasmid ~obtained and identified场alkali splitting and gel electrophoresis. Uposomes~used to transfect UE. RT-PCR based telo- meric repeat amplification protocol(TRAP) assay~used to measure the telomerase activity of endothelia. Results LlE arranged as "cobblestone" and were positive of CD34 labeling. Endothelia transfected场pBABE-HYGR-hTERT(HC ) had an raised absor- bance of 0.889. The shape of growth。~of HC was similar to UE. But the absorbance of MTT test and the amount of HC were prior to UE at every measuring time and the amount of HC increased four times within 8 days ( P

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