期刊论文

中文

中国现代医学杂志

1005-8982

2008

18

13

1845-1848

10.3969/j.issn.1005-8982.2008.13.014

湖南省教育厅资助项目（No:02C391）；

本校为第一机构

目的将人乳头瘤病毒16型(HPV 16)E6蛋白真核表达载体pcDNA3.1(-)/E6转染巨噬细胞,观察E6蛋白瞬时高表达对LPS诱导的THP-1巨噬细胞分泌TNF-α和IL-1β的影响,为进一步研究HPV16E6蛋白的致病机制奠定实验基础。方法将质粒pcDNA3.1(-)/E6通过SuperFectTM脂转染试剂转染巨噬细胞24h后,用LPS刺激巨噬细胞,利用Western-blotting鉴定E6蛋白在巨噬细胞中的表达,ELISA检测pcDNA3.1(-)/E6转染的巨噬细胞不同时间(12、24及48h)培养上清液中TNF-α与IL-1β的含量,统计软件SPSS12.0分析所得数据。结果Western-blotting结果显示pcDNA3.1(-)/E6能在巨噬细胞中表达约18kD的目的蛋白,即E6蛋白;pcDNA3.1(-)/E6+LPS...

[Objective] In order to provide the experimental basis for further studies on the oncogenic mechanism of the E6 protein from human papillomavirus type 16 （HPV16）, the eukaryotic expression vector pcDNA3.1 （-）/E6 was used for the study on the effect of E6 protein to influence the secretory activity of LPS-induced THP-1 macrophages. [Methods] The reconstructed plasmid pcDNA3.1 （-）/E6 was transfected into THP-1 macrophages. The expression of E6 gene was assayed in macrophages lysates by using Western blot analysis and the level of TNF-α or ...