肿瘤抑素30肽的重组表达及抗肿瘤活性研究

Expressionof Recombinant Tumstatin 30 Peptide and Its Antitumor Activity

目的：重组表达肿瘤抑素30肽并研究其抗肿瘤活性。方法设计并合成30肽基因序列,将此序列与融合蛋白表达载体 PTYB21重组,转化到大肠埃希菌 BL-21（DE3）。 IPTG 诱导表达,几丁质柱纯化得到30肽,经 SDS-PAGE 及 Tricine-SDS-PAGE 对纯化产物进行鉴定。利用 MTT 法、吖啶橙/溴化乙锭（AO/ EB）荧光染色法、小鼠 H22腹水转移型肝癌实体瘤抑瘤实验,研究30肽的抗肿瘤活性。结果成功重组并表达肿瘤抑素30肽。体外实验显示,30肽具有抑制 HGC-27胃癌细胞、 HUVEC 人脐静脉细胞增殖和促进这两种细胞凋亡的作用。体内实验显示,30肽对小鼠 H22腹水型肝癌抑瘤率达43.18 ％ 。结论重组的肿瘤抑素30肽具有较强的抗肿瘤活性。

Objective To explore the expression of recombinant tumstatin 30 peptide and its antitumor activity. Methods The 30 peptide sequence was designed, ligated into the fusion protein expression vector PTYB21, and then transformed to E. Coli BL-21 （DE3） . The fusion protein was expressed after induction by IPTG. Tumstatin 30 peptide was purified by using chitin affinity chroma-tography, and verified by SDS-PAGE and Tricine-SDS-PAGE. The antitumor activity of the 30 pep-tide was investigated through MTT assay, AO/ EB fluorescent staining, and the suppression experi-ments in mouse models of H22 ascites liver cancer. Results Recombinant tumstatin 30 peptide was successfully expressed. In vitro experiments showed that the 30 peptide could inhibit the proliferation of gastric cancer cells HGC-27 and human umbilical vein cells （HUVEC） and promote their apopto-sis. The in vivo experiments showed that the inhibition rate of H22 ascites liver tumor by the 30 pep-tide was 43. 18 %. Conclusion Recombinant tumstatin 30 peptide has strong antitumor activity.