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Cloning and biological activity of an anti-tumor peptide of Tumstatin |
| WANG Shujing1, LIU Yan2, LIN Xuesong2, FU Xue2, XU Jianyong2, LIU Xinghan2 |
| 1.Bio-pharmaceutical Key Laboratory of Heilongjiang Province Incubator of State Key Laboratory, Harbin Medical University, Harbin 150086, China; Department of Biochemistry and Molecular Biology, Harbin Medical University, Harbin 150028, China; Institute of Pharmacology, Harbin Commercial University, Harbin 150028, China; 2.Bio-pharmaceutical Key Laboratory of Heilongjiang Province Incubator of State Key Laboratory, Harbin Medical University, Harbin 150086, China; Department of Biochemistry and Molecular Biology, Harbin Medical University, Harbin 150028, China; |
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Abstract To obtain an anti-tumor peptide of Tumstatin and detect its biological activity, the nucleotide sequence encoding 185 203 amino acids (19peptide) of Tumstatin was synthesized and inserted into the fusion protein vector pTYB2. After identification by sequencing and restriction endonucleases, the recombined vector was transformed into BL-21 (DE3) E. coli competent cells. Transformed E. coli BL-21 (DE3) were induced by isopropyl-β-thiogalactopyranoside (IPTG), and then expressed. By 1‚4-dithiothreitol (DTT) reduction, the soluble 19peptide was obtained from a chitin affinity chromatograph. The biological activity of 19peptide was determined by 3-[4‚5-dimethylthiazol-2-y1]-2‚5-dipheny-tetrazolium bromide (MTT) assay, cell growth curve, the effect of the ascitic fluid transfevent H22 hepatoma on mice and via histopathological slices. The purified 19peptide directly inhibited proliferation and migration of murine B16 melanoma cells, SMMC-7721hepatoma carcinoma cells and human umbilical vein endothelial cells (HUVEC). The tumor inhibition rate of mice ascitic fluid transfevent H22 hepatoma was 48.46%. Histopathological slices showed that it could promote tumor tissue necrosis and decrease the density of blood vessels. With higher anti-tumor activity, 19peptide has the potential to become a novel, potent anti-tumor agent.
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Issue Date: 05 September 2007
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