结核分枝杆菌融合蛋白和分泌蛋白对人肝癌细胞HepG-2影响和作用

目的探讨结核分枝杆菌融合蛋白对人肝癌细胞HepG-2的抑制作用。方法构建含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,纯化后复性。分别作用于肝癌细胞HepG-2,MTT法检测细胞生长情况。结果成功纯化并复性三种蛋白。MTT实验结果显示,三种蛋白均对HepG-2细胞的生长具有抑制作用,抑制强度与蛋白终浓度和作用时间相关,但各蛋白的抑制作用无明显差异。结论结核分枝杆菌的部分分泌蛋白对肝癌细胞HepG-2具有抑制作用。

Effects of fusing and excreting protein of mycobacterium tuberculosis on human hepatoma carcinoma cell HepG-2

Objective To investigate the inhibitory effects of mycobacterium tuberculosis（Ag85B-Hsp16.3,Ag85B-ESAT6 and Hsp16.3） on human hepatoma carcinoma cell HepG-2 in vitro.Methods The three recombinant plasmids（pProEXHTa-Ag85B-Hsp16.3,pProEXHTa-Ag85B-ESAT6 and pProEXHTb-Hsp16.3） were established and transferred 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 added into HepG-2 cells respectively and their growth was examined by MTT test assay.Results The proteins were successfully purified and all had moderate inhibitory effects on Hepg-2 cells.The inhibitory effects were dependent on the concentration of the protein as well as the action time.But there was no difference between the inhibitory effects of three proteins.Conclusion Some of the secreted proteins of mycobacterium tuberculosis can inhibit cell growth of hepatoma carcinoma HepG-2.