医学分子生物学杂志
醫學分子生物學雜誌
의학분자생물학잡지
FOREIGN MEDICAL SCIENCES
2015年
2期
75-79
,共5页
赵玲%张忠生%于思茗%刘兴汉
趙玲%張忠生%于思茗%劉興漢
조령%장충생%우사명%류흥한
肿瘤抑素%基因重组%蛋白表达%抗肿瘤活性
腫瘤抑素%基因重組%蛋白錶達%抗腫瘤活性
종류억소%기인중조%단백표체%항종류활성
tumstatin%genetic recombination%protein expression%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。
目的:重組錶達腫瘤抑素30肽併研究其抗腫瘤活性。方法設計併閤成30肽基因序列,將此序列與融閤蛋白錶達載體 PTYB21重組,轉化到大腸埃希菌 BL-21(DE3)。 IPTG 誘導錶達,幾丁質柱純化得到30肽,經 SDS-PAGE 及 Tricine-SDS-PAGE 對純化產物進行鑒定。利用 MTT 法、吖啶橙/溴化乙錠(AO/ EB)熒光染色法、小鼠 H22腹水轉移型肝癌實體瘤抑瘤實驗,研究30肽的抗腫瘤活性。結果成功重組併錶達腫瘤抑素30肽。體外實驗顯示,30肽具有抑製 HGC-27胃癌細胞、 HUVEC 人臍靜脈細胞增殖和促進這兩種細胞凋亡的作用。體內實驗顯示,30肽對小鼠 H22腹水型肝癌抑瘤率達43.18 % 。結論重組的腫瘤抑素30肽具有較彊的抗腫瘤活性。
목적:중조표체종류억소30태병연구기항종류활성。방법설계병합성30태기인서렬,장차서렬여융합단백표체재체 PTYB21중조,전화도대장애희균 BL-21(DE3)。 IPTG 유도표체,궤정질주순화득도30태,경 SDS-PAGE 급 Tricine-SDS-PAGE 대순화산물진행감정。이용 MTT 법、아정등/추화을정(AO/ EB)형광염색법、소서 H22복수전이형간암실체류억류실험,연구30태적항종류활성。결과성공중조병표체종류억소30태。체외실험현시,30태구유억제 HGC-27위암세포、 HUVEC 인제정맥세포증식화촉진저량충세포조망적작용。체내실험현시,30태대소서 H22복수형간암억류솔체43.18 % 。결론중조적종류억소30태구유교강적항종류활성。
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.