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

Ren,Hui; Shao,Xin; Zeng,Liang; Wang,Fa; Huang,Di‑Nan; Hou,Gan

doi:10.3892/mmr.2015.3639

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

Authors:
- Hui Ren
- Xin Shao
- Liang Zeng
- Fa Wang
- Di‑Nan Huang
- Gan Hou
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Affiliations: 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
Published online on: April 17, 2015 https://doi.org/10.3892/mmr.2015.3639
Pages: 2035-2042

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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.

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