大肠埃希菌植物毒素Gelonin核心功能区表达和纯化及抗肿瘤活性研究  

Expression,purification and anti-tumor activity of core functional fragment of plant toxin Gelonin

作　　者：牛作兴[1] 刘爱军[2] 

机构地区：[1]山东省肿瘤医院内科,山东济南250117 [2]解放军总医院病理科,北京100853

出　　处：《中华肿瘤防治杂志》2008年第17期1305-1307,共3页Chinese Journal of Cancer Prevention and Treatment

摘　　要：目的:在大肠埃希菌中表达纯化植物毒素Gelonin核心功能区△Gel(S8-K200),并研究其抗肿瘤活性。方法:以pET30a(+)-Gelonin为模板,采用PCR方法扩增△Gel基因,克隆插入pEASY-B载体,测序鉴定后插入pET30a(+)载体,转化感受态细胞BL21(DE3)pLysS,诱导表达并纯化目的蛋白,并观察其抗肿瘤活性和细胞毒性。结果:△Gel(S8-K200)目的基因的大小约为600bp,SDS-PAGE电泳显示目的蛋白的大小约为27×103,体外实验结果显示,△Gel和Gelonin对肿瘤细胞系HL-60具有明显的杀伤作用,而对正常人胚肺细胞系WI-38毒性极小。结论:Ge-lonin核心功能区△Gel保持了Gelonin毒素蛋白全部的抗肿瘤活性,具有很好的开发前景。OBJECTIVE： To express and purify core functional fragment of plant toxin gelonin and investigate its anti tumor activity. METHODS： The encoding fragment of △Gel（S8-K200） was am plified by PCR with pET30a （＋）-Gelonin as template. The PCR product was ligated with pEASY-B vector and then suhcloned into pET30a（＋ ） vector after conformation by sequencing. The expres- sion vector pET30a（＋）--△Gel was transformed into E. coli BL21 （DE3）pLysS. The target protein fGel was induced and purified so as to investigate its anti-tumor activity and cytotoxicity in vitro. RESULTS： The size of encoding fragment of △Gel（S8 K200）was about 600 bp and SDS-PAGE analysis indicated that the molecular weight of target protein was approximately 27 × 10^3. Both Gelonin and △Gel significantly inhibited the growth of tumor cell line HL 60, and little cytotoxicity to normal cells WI-38 was observed. CONCLUSION ： These results demonstrate that fGel （S8-K200） remain fully anti-tumor activity and provide promise strategies for anti-tumor therapeutics.

关 键 词：细胞毒素类 抗肿瘤药 植物 植物蛋白质/治疗应用 

分 类 号：R91[医药卫生—药学] R730.5