title: Matrix metalloproteinase-mediation of tumor targeting human recombinant tumor necrosis factor-α fusion protein.
abstract: 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 TNF-α (rhTNF-α) fusion protein vectors were constructed. These were subsequently introduced into Escherichia coli. rhTNF-α 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-α fusion protein induces CNE-2Z cells apoptosis. rhTNF-α exhibits high efficacy and tumor cell targeting capability, with low toxicity effects on healthy cells.
authors.affiliation: Department of Basic Medicine, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China.
authors.forename: Hui
authors.initials: H
authors.lastname: Ren
authors.affiliation: Department of Basic Medicine, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China.
authors.forename: Xin
authors.initials: X
authors.lastname: Shao
authors.affiliation: Department of Basic Medicine, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China.
authors.forename: Liang
authors.initials: L
authors.lastname: Zeng
authors.affiliation: Department of Basic Medicine, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China.
authors.forename: Fa
authors.initials: F
authors.lastname: Wang
authors.affiliation: Department of Clinical Biochemistry, Guangdong Medical College, Dongguan, Guangdong 523808, P.R. China.
authors.forename: Di-Nan
authors.initials: DN
authors.lastname: Huang
authors.affiliation: Department of Clinical Biochemistry, Guangdong Medical College, Dongguan, Guangdong 523808, P.R. China.
authors.forename: Gan
authors.initials: G
authors.lastname: Hou
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D020782:Matrix Metalloproteinases / Q000235:genetics / Q000494:pharmacology*; D051379:Mice; D009369:Neoplasms / Q000188:drug therapy; D010957:Plasmids / Q000235:genetics; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000494:pharmacology*
chemical_list: D011993:Recombinant Fusion Proteins; C439524:TNF protein, human; D014409:Tumor Necrosis Factor-alpha; D005982:Glutathione Transferase; D020782:Matrix Metalloproteinases