重组TRAIL_((114-281))片段的表达纯化及抗肿瘤活性分析

Expression and Purification of Recombinant TRAIL_((114-281)) and the Detection of Its Antitumor Activity

本文主要探讨人TRIAL_((114-281))对肠癌和肺癌细胞增殖的影响.利用大肠杆菌原核分泌表达TRAIL_((114-281))片段,Ni-NTA亲和层析柱对表达蛋白进行纯化,纯化的蛋白经SDS-PAGE和Western blotting鉴定,通过CCK-8实验检测TRAIL片段对SW480、DLD1、A549肿瘤细胞增殖的影响.实验结果显示经SDS-PAGE和Western blotting鉴定纯化的蛋白为目的蛋白His-TRAIL_((114-281)),且纯度在90%以上.CCK-8结果显示,在一定浓度下,TRAIL_((114-281))片段能明显抑制SW480、DLD1、A549肿瘤细胞的增殖,抑制率呈浓度依赖性,且对各细胞的抑制作用效果不同.本研究证实TRIAL_((114-281))片段在抗肠癌及非小细胞肺癌细胞方面具有一定的应用前景.

In order to investigate the anti-tumor activity of TRAIL（114-281）. Protein was expressed in Escherichia coli and purified by Ni-NTA chromatography column. SDS-PAGE and Western blotting identify the purified His-TRAIL（114-281）. The changes of cell proliferation were observed by CCK-8 assay. The results show that the target protein was analyzed by SDS-PAGE showing a special band about 20kDa. The target protein was successfully purified by Ni-NTA chromatography. The purity of target protein was above 90%. Western blotting appeared a good antigenicity of the purified protein. The result of CCK-8 assay indicated that the prolifica- tion of SW480, DLD1, 3_549 cells was inhibited significantly. Moreover, this alteration of prolification was in contentration dependent manner. Recombinant TRAIL（114-281）could suppress the prolification of colon and lung cancer , and have some prospect in anti- tumor effects.