携带Survivin-△3(T34A)的减毒鼠伤寒沙门菌疫苗与抗肿瘤效果初探  

Preliminary Study on Oral Attenuated Vaccine Carried Survivin-△3(T34A) Gene and Its Antitumor Effect

作　　者：陈英利[1,2] 祁冬冬[1,3] 李翀[4] 朴金花[1] 张霞[2] 苏晓杰[2] 林雪松[2] 张涛[1] 

机构地区：[1]黑龙江省佳木斯大学生物化学与分子生物学教研室,黑龙江佳木斯154000 [2]哈尔滨医科大学(大庆)医学检验与技术学院 [3]内蒙古林业总医院 [4]宁夏石嘴山市第二人民医院

出　　处：《实用预防医学》2012年第1期8-12,共5页Practical Preventive Medicine

基　　金：黑龙江省卫生厅科技项目(2009-257);黑龙江省研究生创新科研项目专项资金(YJSCX2011-377HLJ);黑龙江省教育厅课题(12511188)

摘　　要：目的探讨携带Survivin-△3(T34A)基因的减毒鼠伤寒沙门菌口服疫苗对乳腺瘤MCF-7注射细胞小鼠的保护作用及其效果。方法人工合成Survivin-△3(T34A)基因,先后将其克隆到真核表达载体pEGFP-N1和pVAX1载体,获得pVAX1-Survivin-△3(T34A)-EGFP真核表达载体,电转化至减毒鼠伤寒沙门菌;小鼠分别口服pVAX1-Survivin-△3(T34A)-EGFP转染沙门菌(实验组)、pVAX1-EGFP转染沙门菌(空质粒组)、沙门菌(对照组)和PBS(空白对照组)后,再皮下注射MCF-7乳腺癌细胞,采用Western-blotting检测Survivin-△3(T34A)蛋白水平,以及测定注射部位瘤块的体积大小。结果成功构建了真核表达载体pVAX1-Survivin-△3(T34A)-EGFP并转化到减毒鼠伤寒沙门菌得到表达;在空质粒组、细菌组、空白对照组和实验组小鼠中,肿瘤瘤块平均直径分别为(25.813±0.108)mm、(25.627±0.117)mm、(26.257±0.213)mm、(9.124±0.086)mm;实验组与其他组相比,差异有统计学意义(P<0.05),且能在血清中检测到Survivin-△3(T34A)-EGFP融合蛋白的存在。结论小鼠口服携带Survivin-△3(T34A)真核质粒的沙门菌后,能够抑制MCF-7乳腺瘤细胞的增殖和扩散。Objective To explore the protection role and the effect of oral attenuated Salmonella typhimuriurn vaccine carried Survivin - A3 （T34A） gene against MCF - 7 breast tumor cells in mice. Methods Artificial synthetic Survivin - A3 （T34A） gene was cloned into the eukaryotic expression vector pEGFP - N1 and pVAX1 orderly, the pVAX1 - Survivin A3 （T34A） - EGFP eukaryotic expression vector was obtained and then transformed into attenuated Salmonella typhimurium. The mice were divided into four groups which took orally pVAX1 - Survivina3 （T34A） - EGFP transfection Salmonella （test group）, pVAX1- EGFP transfection Salmonella （empty plasmid group）, Salmonella （control group） and PBS （blank control group）, respectively, and then the mice of each group were injected subcutaneously with MCF- 7 breast tumor cells. The levels of Survivin-△3 （T34A） protein were detected by Western- blotting, and the tumor sizes of injection sites of mice were mea- sured. Results The eukaryotic expression vector pVAXl - Survivin - 3 （T34A） - EGFP was constructed successfully and then transformed into the attenuated Salmonella typhimurium. The mean diameters of the tumors in empty plasmid group, bacteria group, blank control group and test group were （25. 813±0. 108） rnm, （25. 627±0. 117） mm, （26. 257±0. 213） mm, and （9. 124±0. 086） mm, respectively. There were statistically significant differences between test group and the other groups （ P 〈 0.05 ） ; moreover, Survivin - 3 （T34A） - EGFP fusion protein was detected in the sera of the mice in test group. Conclusions After oral administration of the Salmonella carried the Survivin- A3 （T34A） eukaryotic expression plasmids, the prolif-eration and diffusion of MCF- 7 breast tumor cells in mice can be inhibited.

关 键 词：存活素 减毒鼠伤寒沙门菌 Survivin—△3 Survivin—△3(T34A) PVAX1 

分 类 号：R730.59[医药卫生—肿瘤]