The aim of the present study was to use genetic engineering in order to establish an efficient tumor necrosis factor (TNF)α fusion protein with low toxicity, which may be used to target tumors. Four types of matrix metalloproteinase (MMP)mediated tumor targeting human recombinant
TNFα (rhTNFα) fusion protein vectors were constructed. These were subsequently introduced into Escherichia coli. rhTNFα fusion protein with a glutathione Stransferase (GST)tag was purified using GST resin affinity chromatography, and GSTtags 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 GSTtagged 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, CNE2Z, which secretes high levels
of MMP1. In conclusion, the results of the present study suggested that MMPmediated rhTNFα fusion protein induces CNE2Z cells apoptosis. rhTNFα exhibits high efficacy and tumor cell targeting capability, with low toxicity effects on healthy cells.
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Document Type: Research Article
Department of Basic Medicine, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China
Department of Clinical Biochemistry, Guangdong Medical College, Dongguan, Guangdong 523808, P.R. China
Publication date: August 1, 2015
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Molecular Medicine Reports is a monthly, peer-reviewed journal available in print and online, that includes studies devoted to molecular medicine, underscoring aspects including pharmacology, pathology, genetics, neurosciences, infectious diseases, molecular cardiology and molecular surgery. In vitro and in vivo studies of experimental model systems pertaining to the mechanisms of a variety of diseases offer researchers the necessary tools and knowledge with which to aid the diagnosis and treatment of human diseases.
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