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| Expression and Purification of Recombinant Human Glutathione Peroxidase 7 and the Detection of Its Tumour Suppressive Effect |
| CHEN Xin-lin, QIAN Xiu-ping, ZHU Jian-wei, GE Mei, CHEN Dai-jie, Mao Wen-wei |
| Laboratory of Microbial and Biochemical Pharmaceuticals, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China |
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Abstract Recent study reported that reconstitution expression of GPx7 in tumour cell led to great suppressive effect on tumour growth in mouse xenograft model. Consequently, we carried out an administration of GPx7 through subcutaneous injection to investigate effect of GPx7 on tumour growth in mouse xenograft model. The coding domain sequence of human GPx7 without putative signal peptide (amino acid 1-19) was amplified from previously constructed plasmid pcDNA3.1(-)-hGPx7 and was subcloned into prokaryotic expression pET24A vector. The GPx7 purification process included two chromatography steps in which cation exchange and size exclusion chromatography was adopted. The resulting pure product showed weak and limited GPx activity in vivo. Purified GPx7 was injected subcutaneously through scruff of the neck of xenograft mouse model. The result showed that after GPx7 injection for 14 consecutive days, GPx7-injecting group showed to have a slower tumour growth compared with control (P<0.05) and the inhibition rate is 29.26%. The result indicated that two purification steps which started from cation exchanger and followed by size exclusion are practical and effective, meanwhile, GPx7 also showed to have tumour suppressive effect in vivo.
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Received: 20 May 2014
Published: 25 July 2014
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