关键词: 细胞凋亡, 癌, 非小细胞肺, 顺铂

Abstract: Objective To construct prokaryotic expression plasmid of human recom binant soluble TNF-related apoptosis inducing ligand(rsTRAIL), then to investig ate the effects of rsTRAIL on apoptosis in non-small cell lung carcinoma（NSCLC）. Methods The encoding sequence for rsTRAIL was amplified with RT-PCR and cloned into PQE₃0 vector to establish the prokaryotic expression system. The competent cells of host strain of M₁5 were transformed by the recombinant plasmid. The expression of the target protein was induced with IPTG and purified by Ni²⁺ -NTA agarose column. rsTRAIL was added in the media of A549 and H460^wt cells, then the viability was examined by MTT assay. Apoptosis of H460^wt cells was observed under fluorescence microscopy. The apoptotic rate of tumor cells was examined by FACS. Results The cloned fragment of rsTRAIL was 100% consistent with that reported in GenBank. The expressed protein with molecular weight of 21 000 in SDS-PAGE as expected was obtained and recognized by a commercial McAb. The apoptotic rate of H460^wt cells after treated with rsTRAIL for 24 h was 43.2%. Cislatin enhanced the effect of rsTRAIL on A549 cells. Conclusion The rsTRAIL is obtained after Ni affinity chromatograph. rsTRAIL has a strong cytotoxic activity against NSCLC and cisplatin may enhance the antit umor effect of rsTRAIL .

Key words: apoptosis, carcinoma, non-small cell lung, cisplatin