Matrix metalloproteinase-mediation of tumor targeting human recombinant tumor necrosis factor-α fusion protein

The aim of the present study was to use genetic engineering in order to establish an efficient tumor necrosis factor (TNF)-alpha fusion protein with low toxicity, which may be used to target tumors. Four types of matrix metalloproteinase (MMP)-mediated tumor targeting human recombinant TNF-alpha (rhTNF-alpha) fusion protein vectors were constructed. These were subsequently introduced into Escherichia coli. rhTNF-alpha fusion protein with a glutathione S-transferase (GST)-tag was purified using GST resin affinity chromatography, and GST-tags were digested using factor Xa. The cytotoxic effects of the fusion protein on L929 cells were determined using MTT assays. At a concentration of 1 pM, the GST-tagged fusion protein exerted no cytotoxic effects on the cells, compared with the negative control cells (P=0.975>0.05). However, at a concentration of 1000 pM, the deblocking fusion protein exerted greater cytotoxic effects on L929 cells, compared with positive control cells (P<0.05). Treatment with the fusion protein also induced cell apoptosis in the nasopharyngeal cancer cell line, CNE-2Z, which secretes high levels of MMP-1. In conclusion, the results of the present study suggested that MMP-mediated rhTNF-alpha fusion protein induces CNE-2Z cells apoptosis. rhTNF-alpha exhibits high efficacy and tumor cell targeting capability, with low toxicity effects on healthy cells.