5HRE增强子和hTERT启动子联合调控CDX2基因对裸鼠移植瘤的抑制作用  

Effects of regulation of CDX2 gene by 5HRE enhancer and hTERT promoter on growth of colorectal transplanted tumors in nude mice

作　　者：祁杰[1] 郑见宝[2] 孙学军[2] 王孝珑 余钧辉[2] 吴云桦 高琪[2] 王恺[2] 贺赛 QI Jie ZHENG Jianbao SUN Xuejun WANG Xiaolong YU Junhui WU Yunhua GAO Qi WANG Kai HE Sai(Department of Vascular Disease, Shaanxi Provincial Hospital, Xi＇ an 710068, China Department of General Surgery, First Affiliated Hospital of Xi＇ an Jiaotong University Department of Surgical Oncology, First Affiliated Hospital of Xi＇ an Jiaotong University Department of Breast Cancer, Shaanxi Provincial Tumor Hospital)

机构地区：[1]陕西省人民医院心内二科,西安710068 [2]西安交通大学第一附属医院普通外科 [3]西安交通大学第二附属医院肿瘤外科 [4]陕西省肿瘤医院乳腺病院

出　　处：《山西医科大学学报》2017年第3期246-250,共5页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(81101874;81172362);陕西省科学技术研究发展计划项目(2016SF-015);陕西省科技统筹创新工程计划项目(2013KTCQ03-08)

摘　　要：目的探讨5HRE增强子和hTERT启动子联合调控CDX2基因对结肠癌LoVo细胞裸鼠皮下移植瘤的抑制作用,为结肠癌生物治疗提供实验依据。方法将复苏前期筛选的稳定表达pLVX-hTERTp-CDX2-3FLAG(hC)的LoVo细胞(hC/LoVo)、空LoVo细胞、稳定表达pLVX-5HRE-hTERTp-3FLAG(5Hh)的LoVo细胞(5Hh/LoVo)及转染pLVX-5HRE-hTERTp-CDX2-3FLAG(5HhC)的LoVo细胞(5HhC/LoVo),接种各组细胞制备裸鼠皮下移植瘤模型,观察构建的载体对LoVo细胞移植瘤生长的影响;免疫组化法观察移植瘤CDX2和Ki-67的表达情况。结果成功复苏转染5HhC的LoVo细胞及各组对照LoVo细胞。各组裸鼠皮下接种肿瘤细胞后均成功致瘤,成瘤率为100%;5HhC组种植瘤生长速度显著低于其他组(P<0.05);终点时间第18天时,5HhC组裸鼠皮下种植瘤的体积明显小于其他各组(P<0.05);5HhC组种植瘤质量为(0.20±0.16)g,明显低于hC/LoVo组(0.34±0.21)g,空LoVo细胞组(0.61±0.19)g及5Hh/LoVo组(0.55±0.27)g(P<0.05);5HhC组种植瘤中CDX2表达为(87±7)%,明显低于hC/LoVo组(55±4)%,空LoVo细胞组(15±3)%及5Hh/LoVo组(17±5)%(P<0.05);5HhC组种植瘤中Ki-67表达为(7±7)%,明显低于hC/LoVo组(13±2)%,空LoVo细胞组(38±5)%及5Hh/LoVo组(85±9)%(P<0.05)。结论构建的双靶向调控载体5HhC可以抑制结肠癌LoVo细胞裸鼠移植瘤的生长。Objective To verify the effect of pLVX-5HRE-hTERTp-CDX2-3FLAG（5HhC）containing the tumor suppressor gene CDX2 regulated by the hypoxia-induced enhancer（HRE） and the hTERT promoter on growth of colorectal transplanted tumors in mude mice. Methods The LoVo cells with stable expression of pLVX-hTERTp-CDX2-3FLAG （hC）, pLVX-SHRE-hTERTp-3FLAG （5Hh）and pLVX-SHRE-hTERTp-CDX2-3FLAG（5HhC）were obtained by transfection technique, named as hC/LoVo,5Hh/LoVo and 5HhC/Lo- Vo. A xenograft tumor model was established and tumor growth was observed. The expression of CDX2 and Ki-67 protein in tumor tissues were detected by imm^nohistochemistry. Results The xenograft tumor models of human colorectal carcinoma cells in hC/LoVo, LoVo ,5Hh/LoVo and 5HhC/LoVo groups were established successfully. The tumor volume in 5HhC/EoVo group was significantly smal- ler than in the other three groups at every time point （P 〈 0.05 ）. At the termination of observation, the tumor weight in 5HhC/Lovo group was significantly smaller than in hC/LoVo, LoVo and 5 Hh/LoVo groups respectively [ （0.20± 0.16 ） g vs （ 0. 34± 0.21 ） g, （0. 61 ±0. 19）g, （0. 55 ± 0. 27 ）g, P 〈 0.05 ]. The expression of CDX2 protein by immunohistochemistry in tumor tissues in 5HhC/LoVo group [ （ 87 ±7 ） % ] were higher than that of hC/LoVo [ （ 55 ±4 ） % ], LoVo [ （ 15± 3 ） % ] and 5Hh/LoVo [ （ 17 ± 5 ） % ], respectively （P 〈 0. 05 ）, while the expression of Ki-67 protein in tumor tissues in 5 HhC/LoVo group was lower than in the other three groups [ （7 ±7）%vs （13 ±2）%,LoVo（38 ±5）% and 5Hh/LoVo（85 ±9）% ,P 〈0.05]. Conclusion The gene therapy vector 5HhC may in- duce the growth of colorectal transplanted tumors in mude mice.

关 键 词：基因治疗载体 结肠癌 裸鼠 移植瘤 CDX2 Ki-67 

分 类 号：R735.35[医药卫生—肿瘤]