小分子肽-肿瘤抑素19肽纯化及抗肿瘤活性研究

Study on Anti-tumor Activity and Purification of Tumor Chalone 19 Peptide

目的:利用已构建好的肿瘤抑素抗肿瘤活性肽-19肽高效表达基因重组菌纯化19肽,并进行抗肿瘤活性检测。方法:经几丁质亲和层析柱纯化出19肽,SDS-PAGE聚丙烯酰胺电泳后用Bradford法测其浓度,进行体内活性检测。结果:19肽可促进H22小鼠肝癌细胞死亡,血管数量减少,抑瘤率达48．46%。结论:得到了具有抗肿瘤活性的肿瘤抑素相关肽—19肽,为肿瘤抑素的作用机制研究和肿瘤的临床治疗奠定了基础。

Objective： To construct anti-tumor peptide of tumor chalone-19 peptide genetic engineering bacteria and tc identify its biological activity, Methods： The base sequence of 19 pepfide was designed,artificially synthesized and inserted into the fusion protein vector pTYB2.After sequencing and restriction, the recombinant 19 peptide was transformed and expressed in E.coli BL-21 （DEB） by means of fusion protein. By DTF reduction the soluble 19 pepfide was obtained from chitin affinity chromatograph. By such experiment as MT-F assay, cell growth curve, the effect of the ascitic fluid transfevent H22 hepatoma of mice and histopathological slice, the biological activity of 19 pepfide was studied. Results： The optimal expression condition of recombinant 19 peptide was 28℃,0.1mM IPTG, 6h .The purified 19 peptide direcdy inhibited proliferation and migration of hepatoma carcinoma cell. The tumor inhibition rate of ascitic fluid transfevent H22 hepatoma of mice was 48.46%. Histopathological slice showed it could promote tumor tissue necrosis and decrease the number of blood vessel. Conclusions： We can obtain recombinant 19 peptide of tumor chalone with anti-tumor activity, which lay a foundation for its mechanism of action research and clinical tumor therapy.