The regulation of the TACE pro-domain to tumor necrosis factor-? converting enzyme activity / 中国病理生理杂志

ABSTRACT

AIM:To study functions of pro-domain in tumor necrosis factor-? converting enzyme (TACE) maturation and to develop an approach to interfere with inflammation processes. METHODS: The plasmid pIRES2-EGFP was used to generate the expression plasmids constructed with the signal peptide and pro-domain, full length or without the pro-domain(designated as pIRES2-EGFP/T648, pIRES2-EGFP/T2472, pIRES2-EGFP/T57-T1824,respectively). The plasmids were used to transfect the U937 cells.After the transfected cells were stimulated by LPS, the effect of expression plasmids on TNF-? secretion was detected by ELISA and flow cytometry(FCM).RESULTS: The plasmid pIRES2-EGFP/T648 inhibited 61.09% TNF-? secretion and mediated the accumulation of TNF-? on the surface of U937 cells.The sTNF-? secretion and the level of the mTNF-? in the plasmid pIRES2-EGFP/T57-T1824 transfected cells had no significant deviation compared with the pIRES2-EGFP transfected cells. The sTNF-? secretion of the cells transfected by the plasmid pIRES2-EGFP/T2472 increased and the level of the mTNF-? decreased.CONCLUSION: The pro-domain has dual effects in TACE maturation and might be applied to molecular design of anti-inflammation drug.