| 杨巍1 师长宏2△ 张彩勤2 赵勇2 赵善民2 赵佐庆1.结核分枝杆菌融合蛋白Ag85B-Hsp16.3、Ag85B-ESAT6 和分泌蛋白
Hsp16.3 体外对人肝癌细胞HepG-2 的作用*[J].,2011,11(10):1801-1804 |
| 结核分枝杆菌融合蛋白Ag85B-Hsp16.3、Ag85B-ESAT6 和分泌蛋白
Hsp16.3 体外对人肝癌细胞HepG-2 的作用* |
| Effect of Ag85B-Hsp16.3 Ag85B-ESAT6 and Hsp16.3 of MycobacteriumTuberculosis on Human Hepatoma Carcinoma Cell HepG-2 in Vitro |
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| DOI： |
| 中文关键词: 结核分枝杆菌；Ag85B；ESAT6；Hsp16.3；肝癌 HepG-2 |
| 英文关键词: Mycobacterium tuberculosis Ag85B ESAT6 Hsp16.3 HepG-2 |
| 基金项目:国家“863”专项课题（2007AA02Z473），国家自然科学基金（NO.30972767），陕西省自然科学基金项目（SJ08C203）资助 |
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| 中文摘要: |
| 目的：探讨结核分枝杆菌融合蛋白Ag85B-Hsp16.3、Ag85B-ESAT6 及分泌蛋白Hsp16.3 对人肝癌细胞HepG-2 的作用。方
法：将已构建的含3 种目的基因的表达载体pProEXHTa-Ag85B-Hsp16.3、pProEXHTa-Ag85B-ESAT6 和pProEXHTb-Hsp16.3，分
别转入宿主菌E.coli DH5α 中，诱导表达后分别获得Ag85B-Hsp16.3、Ag85B-ESAT6 和Hsp16.3 三种蛋白，采用Ni2+ 亲和层析柱
进行纯化，并用透析方法进行目的蛋白的复性。复性的蛋白按照不同浓度和作用时间分别与肝癌细胞HepG-2 反应，用MTT 法检
测细胞生长情况。结果：三种蛋白被成功纯化并复性。MTT 数据统计分析显示，终浓度10μg/ml 的三种蛋白对HepG-2 细胞生长
没有明显作用，当三种蛋白的终浓度分别为20、40、80μg/ml 时均能够抑制HepG-2 细胞的生长，并且抑制作用随着蛋白终浓度的
增大以及作用时间的延长而增强。不同类别的蛋白抑制作用没有明显差别。结论：结核分枝杆菌的部分分泌蛋白能够抑制肝癌细
胞HepG-2 的生长。 |
| 英文摘要: |
| Objective: To investigate the effect of Ag85B-Hsp16.3, Ag85B-ESAT6 and Hsp16.3 of Mycobacterium tuberculosis
(MTB) on HepG-2 in vitro. Methods：The three recombinant plasmids (pProEXHTa-Ag85B-Hsp16.3, pProEXHTa-Ag85B-ESAT6 and
pProEXHTb-Hsp16.3) were transfered into E.coli DH5α respectively. The three proteins were induced by IPTG and purified by Ni-NTA
purification system under denaturing condition. Following renaturation by dialysis and filtration, the three proteins were respectively
added into HepG-2 cells of various concent ration (10μg/ml,20μg/ml,40μg/ml and 80μg/ml). The cells were incubated for 24h or 48h,
and then the inhibition rate was examined by MTT test assay. Results：The proteins were successfully purified and all had a moderate
killing effect on Hepg-2 cells. The effect was dependent on the concent ration of the protein as well as the action time.But there was no
statistical difference between different proteins. Conclusion：Some of the secreted proteins of Mycobacterium tuberculosis can inhibit cell
growth of liver cancer cell HepG-2. |
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