裸鼠皮下移植人神经母细胞瘤模型的建立和HIF-1α仅及其相关蛋白的表达  

The HIF-1α and its related proteins＇ expression in an athymic mouse human neuroblastoma xenograft model

作　　者：王家祥[1] 孟庆磊[1] 张大[1] 穆鑫[1] 张国锋[1] 

机构地区：[1]郑州大学第一附属医院小儿外科,450052

出　　处：《中华小儿外科杂志》2011年第8期604-609,共6页Chinese Journal of Pediatric Surgery

基　　金：河南省科技厅杰出人才基金资助项目（编号：94200510015）

摘　　要：目的探讨应用人神经母细胞瘤细胞株建立裸鼠皮下神经母细胞瘤模型，测定缺氧诱导因子（hypoxia-inducible factor-1，HIF-1α）及其相关蛋白在该模型中的表达，了解HIF-1α及其相关蛋白在神经母细胞瘤发生发展中的作用，为神经母细胞瘤（neuroblastoma NB）的发病机制研究和诊治提供新的思路与方法。方法应用KP-N-NS人肾上腺神经母细胞瘤细胞株建立模型。采用S-P免疫组织化学方法检测裸小鼠皮下移植瘤、手术切除的神经母细胞瘤肿瘤标本、正常人肾上腺髓质组织中HIF-1α、VEGF及TGF-α的表达情况。结合免疫组化结果对HIF-1α阳性组与HID-1α阴性组裸鼠的体重变化、瘤体重量、存活天数进行比较，并对实验结果进行统计学处理。结果4周后解剖结果证实成模率80％；HIF-1α、VEGF及TGF-α“在肿瘤组织中的阳性表达率显著高于正常组织（P〈0．01），肿瘤组与阳性对照组HIF-1α、VEGF、TGF-α三种蛋白的阳性染色的积分光密度（integrating optical density，10D）值明显高于阴性对照组（P〈0．01）。HIF-1α在肿瘤组与阳性对照组中的表达与VEGF的表达成正相关（P〈0．05），与TGF-α的表达无明显相关性（P〉0．05）；HIF-1α阳性组与HIF-1α阴性组体重变化、瘤体重量及存活天数比较，差异有统计学意义（P〈O．01）；HIF-1α阳性组较HIF-1α阴性组裸鼠生存时间短（P〈0．05）。结论该模型用于研究人类神经母细胞瘤，造模时间短，成瘤率高，生物学行为与人类神经母细胞瘤接近。HIF-1α、VEGF及TGF-1α蛋白的阳性表达与神经母细胞瘤的浸润及转移有关。HIF-1α与VEGF在神经母细胞瘤的发生发展过程中可能存在一定的协同作用。HIF-1α的高表达是预后不良的一个重要指标。Objective To study the HIF-1αand its related proteins＇ expression in an athymicmouse human neuroblastoma xenograh model. Methods The was established by subcutaneous injection of human adrenal neuroblastoma cells. The expressions of HIF-la, VEGF and TGF-α were detected by S-P immunohistochemical techniques. The tumor group was divided into HIF-1α positive group and the HIF-la negative group according to the immunohistochemistry results. The body weight, tumor weight and survival days of the HIF-lapositive mice and HIF-lanegative mice were statistically analyzed. Results Four weeks later, significant neuroblastoma growth occurred in 40 mice （80%）. The positive HIF-1α, VEGF and TGF-α rate of the tumor group was （73.70 ± 10. 68） %, （69. 80 ± 9. 91） %, and （71.43 ± 8. 52） %, respectively. It is significantly increased compared with the control group （P〈0. 01）, which were （9. 67 ± 4. 53） %, （6. 80 ±.40） %, and （6. 50± 4. 44） %, respective-ly. The integrated optical density （IOD） of HIF-1α, VEGF and TGF-a was 141.97 ± 7. 98,151.85 ±14. 35, and 139. 94 ± 4. 50 in tumor group, and 141.34 ± 6. 44, 144. 06 ± 8.51 and 149.00 ±13.63 in positive control group. They were significantly higher than those of the negative control group （P〈 0. 01）. In the tumor group and positive control group, the expression of HIF-1α was positively correla- ted with the expression of VEGF （%2 = 7. 778, r = 0. 504, P〈0. 01）. However, no correlation between the expression of HIF-1α and TGF-a was found（X^2 = 0. 208,r= 0. 934,p〉0. 05）. The body weight, tumor weight and survival days of HIF-1α positive mice were significantly different with those of HIF- 1α negative mice （P〈0. 01）. The mice with positive HIF-1α expression had shorter survival time than the mice with negative HIF-1α （X^2 = 19. 013,P〈0. 05）. Conclusions The athymic mouse human neuroblastoma xenograft model is a good model to research the biology behavior of human neuroblastoma. The overexpressio

关 键 词：缺氧诱导因子1 神经母细胞瘤 免疫组织化学 小鼠 裸 

分 类 号：R739.4[医药卫生—肿瘤]