| 端粒酶催化亚单位转染血管内皮细胞的实验研究 |
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| 引用本文： | 代晓明,李龙江,温玉明,王昌美,刘华,刘坤,李胜富. 端粒酶催化亚单位转染血管内皮细胞的实验研究[J]. 华西口腔医学杂志, 2004, 22(5): 373-375 |
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| 作者姓名： | 代晓明 李龙江 温玉明 王昌美 刘华 刘坤 李胜富 |
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| 作者单位： | 1.口腔生物医学I程教育部重点实验室,四川大学; 2.四川大学华西医院移植免疫室,四川成都610041 |
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| 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<0.05). UE与HC的细胞生长曲线形态相似,8d内HC数量增长了约4倍。在各个时间点HC 较UE细胞数多(P < 0.05)。结论pBABE-HYGRO-hTERT转染内皮细胞后,提高了内皮细胞端粒酶的活性和增殖能力。
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| 关 键 词： | 血管内皮细胞 端粒酶催化亚单位 转染 |
| 收稿时间： | 2004-10-25 |
| 修稿时间： | 2004-10-25 |
Studies on the transfection of umbilical endothelia with catalytic subunit of telomerase] |
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| Xiao-ming Dai,Long-jiang Li,Yu-ming Wen,Chang-mei Wang,Hua Liu,Kun Liu,Sheng-fu Li. Studies on the transfection of umbilical endothelia with catalytic subunit of telomerase][J]. West China journal of stomatology, 2004, 22(5): 373-375 |
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| Authors: | Xiao-ming Dai Long-jiang Li Yu-ming Wen Chang-mei Wang Hua Liu Kun Liu Sheng-fu Li |
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| Affiliation: | Key Laboratory of Oral Biomedical Engineering Ministry of Education, Sichuan University, Chengdu 610041, China. |
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| Abstract: | OBJECTIVE: To investigate the variety of proliferating ability of umbilical endothelia (UE) transfected by plasmid pBABE-HYGR-hTERT. METHODS: UE was identified from two aspects: morphology and CD34 labeling technique. The plasmid was obtained and identified by alkali splitting and gel electrophoresis. Liposomes were used to transfect UE. RT-PCR based telomeric repeat amplification protocol (TRAP) assay was used to measure the telomerase activity of endothelia. RESULTS: UE arranged as "cobblestone" and were positive of CD34 labeling. Endothelia transfected by pBABE-HYGR-hTERT(HC) had an raised absorbance of 0.889. The shape of growth curve 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 < 0.05). CONCLUSION: The transfection of pBABE-HYGRO-hTERT had greatly improved the proliferating abilities and activated the telomerase of UE. |
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| Keywords: | vascular endothelium catalytic subunit of telemerase |
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