Antitumor Activity of Recombinant Soluble Human TRAIL

As a potential antitumor protein, tumor necrosis factor-related apoptosis-inducing ligand(TRAIL) has drawn considerable attention. The coding region human soluble TRAIL gene from LPS activated healthy human peripheral lymphocytes (PBL) was cloned using RT-PCR and constructed 6 His-tag vector pET30a-TRAIL(114~281)and pET30a-TRAIL(120~281).After DNA sequencing, The expression of protein was induced by 0.2 mmol/L IPTG and low temperature(4 ℃). Soluble protein was purified by Ni-NTA chromatography column. Purified protein was digested with Enterkinase to cut off 6×His-tag and was purified by Gel filtration chromatography, showing a peak at 6.0×10~4 or so. Results of SDS-PAGE proved that the 6.0×10~4 protein was TRAIL trimer. The MTT assay showed that TRAIL(114~281) and TRAIL(120~281) could inhibit the growth of HeLa cell line, at the same time, protein B could inhibit the growth of some types of tumor cell and good relationship of concentration-effect and time\|effect.