腺病毒介导生长抑素2型受体对裸鼠人胰腺癌移植瘤抑制作用的研究  

Adenoviral-mediated somatostatin receptor subtype 2 gene transfer inhibits pancreatic cancer xenografts.

作　　者：徐彬[1] 徐浩[2] 李月琴[1] 曹忠健[1] 张欣[1] 周天鸿[1] 

机构地区：[1]暨南大学生物工程学系,广东广州510632 [2]暨南大学医学院第一附属医院核医学科,广东广州510632

出　　处：《中国实用内科杂志》2007年第8期593-596,共4页Chinese Journal of Practical Internal Medicine

基　　金：广东省重大科技计划项目(2003A3080501)

摘　　要：目的构建表达生长抑素2型受体(SSTR2)的重组腺病毒,探讨其对裸鼠人胰腺癌移植瘤生长的影响及机制。方法2004-06-01—2005-12-10,在暨南大学生物工程学系采用位点特异性重组方法构建和包装携带人生长抑素2型受体和报告基因LacZ的重组腺病毒Ad-SSTR2和Ad-LacZ。建立裸鼠人胰腺癌移植瘤模型,分别于瘤内注射生理盐水(阴性对照组)、Ad-LacZ(报告基因对照组)和Ad-SSTR2(实验组),观察肿瘤大小和重量变化。通过逆转录-聚合酶链反应(RT-PCR)和免疫印迹试验(Western blot)鉴定SSTR2在裸鼠肿瘤组织中的表达,Western blot测定信号传导通路蛋白ERK2和ras的表达情况。结果获得滴度分别为6.0×1012pfu/L和6.5×1012pfu/L的重组腺病毒Ad-SSTR2和Ad-LacZ。Ad-SSTR2转染裸鼠胰腺癌移植瘤后SSTR2 mRNA和蛋白都得到有效表达,实验组对移植瘤生长具有明显的抑制作用,抑制率为48.2%。与阴性对照组和报告基因对照组相比,实验组ERK2和ras蛋白的表达量明显减少(P<0.01)。结论腺病毒介导的生长抑素2型受体对裸鼠人胰腺癌移植瘤的生长具有抑制作用,其作用机制可能与信号传导通路蛋白ERK2和ras的表达下调有关,提示其有可能成为胰腺癌基因治疗的一种有效方法。Objective To construct recombinant adenovirus carrying the gene of human somatostatin receptor subtype 2 （SSTR2） ,to investigate its effect on growth of human pancreatic cancer xenografts in nude mice,and to further elucidate the underlying mechanisms. Methods The recombinant adenovirus coding for human SSTR2 （ Ad-SSTR2 ） and reporter gene LacZ（Ad-LacZ） were generated by site-specific recombination from Jun 1,2004 to Dec 10,2005. Human pancreatic cancer cell llne was implanted subcutaneously into nude mice;normal saline（ control group） , Ad-LacZ（ reporter gene con- trol group）and Ad-SSTR2 （experimental group）were injected into pancreatic cancer xenografts, respectively. The tumor volume and weight were measured. RT-PCR and Western blot were used to determine the expression of SSTR2 after pancreatic cancer xenografts were infected with Ad-SSTR2. The expression level of ERK2 and ras proteins were assessed by Western blot. Results The virus titer of Ad-SSTR2 and Ad-LacZ was 6. 0 × 10^12pfu/L and 6. 5 × 10^12pfu/L,respectively. SSTR2 mRNA and protein were detected after Ad-SSTR2 infected pancreatic cancer xenografts. Growth of pancreatic cancer was significantly inhibited in the experimental group as compared with the control group. The growth inhibitory rate was 48.2% （ P 〈 0.01 ）. The significant decrease in expression of ERK2 and ras proteins was detected in the experimental group compared with the reporter gene control group and control group. Conclusion The recombinant adenovirus coding for SSTR2 has inhibitory effect on growth of human pancreatic cancer xenografts ; the mechanisms are associated with down-regulation of the expression of ERK2 and ras proteins. It might be a useful treatment approach to gene therapy of human pancreatic cancer.

关 键 词：受体 生长抑素 胰腺肿瘤 腺病毒 基因治疗 

分 类 号：R5[医药卫生—内科学]