骨髓基质干细胞在Sprague-Dawley大鼠颅内移植致肿瘤形成3例报告  

Tumorigenesis of bone marrow stromal cells transplanted into the brain of Sprague-Dawley rats:report of three cases

作　　者：刘振林[1] 王亮[1] 刘晓智[1] 刘洪良[1] 宋军[2] 苏治国[1] 宋迎[2] 李金艳[2] 姜忠敏[3] 

机构地区：[1]天津市第五中心医院神经外科,天津300450 [2]天津市第五中心医院神经内科,天津300450 [3]天津市第五中心医院病理科,天津300450

出　　处：《肿瘤》2011年第10期881-885,共5页Tumor

基　　金：天津市应用基础及前沿技术研究计划资助项目(编号:09JCYBJC09500);国家自然科学基金资助项目(青年科学基金项目)(编号:81000901)

摘　　要：目的:探讨骨髓基质干细胞(bone marrow stromal cells,BMSCs)颅内移植后潜在的恶变致瘤的可能性及机制。方法:选取在20只Sprague-Dawley大鼠体内移植BMSCs实验中发现的3只对侧肢体偏瘫症状逐渐加重的实验大鼠,采用MRI检查其颅内是否发生占位性改变,观察并记录此3只大鼠的生存期;获取死亡大鼠的含瘤脑组织,采用端粒扩增(telemere repeat amplifi cation protocol,TRAP)-PCR-酶联免疫(enzyme-linked immunosorbent assay,ELISA)法检测细胞端粒酶活性,免疫组织化学法检测含瘤脑组织中细胞核增殖抗原(proliferating cell nuclear antigen,PCNA)、巢蛋白(nestin)和胶质纤维酸性蛋白(glial fi brillary acidic protein,GFAP)的表达,蛋白质印迹法检测表皮生长因子受体(epidermal growth factor receptor,EGFR)、血管内皮生长因子(vascular endothelial growth factor,VEGF)和基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)的相对表达量。以无瘤正常大鼠脑组织为对照组。结果:20只BMSCs移植大鼠中共有3只实验鼠出现左侧肢体偏瘫且呈进行性加重,3只大鼠的生存期分别为129、174和187d;TRAP-PCR-ELISA结果显示,3只含瘤组织中细胞端粒酶活性均高于无瘤组;免疫组织化学结果显示,3只大鼠含瘤组织中PCNA和nestin的蛋白表达水平均高于无瘤组,GFAP表达水平则低于无瘤组;蛋白印迹法检测结果提示,与无瘤组织比较,3例肿瘤组织中有1只EGFR表达增高,2只VEGF表达增高,而MMP-9表达水平差异均无统计学意义。结论:长期体外培养的BMSCs可出现端粒酶活性和细胞增殖活力增强,以及部分癌基因激活等致瘤表型,提示干细胞体内移植存在潜在的成瘤可能性。Objective： To investigate the possible oncogenicity of bone marrow stromal stem cells （BMSCs） transplanted into brain and its underlying mechanism. Methods： The contralateral hemiplegia became gradually worse in three Sprague-Dawley rats among twenty rats after intracerebral transplantation with BMSCs. The intracerebral space occupying lesions were examined by MRI, and the life span of these rats was observed and recorded. The tumor tissues were obtained after the three rats were sacrificed. The telomerase activity was detected by telemeric repeat amplification protocol （TRAP）- PCR-enzyme-linked immunosorbent assay （ELISA）. The expressions of proliferating cell nuclear antigen （PCNA）, nestin and glial fibrillary acidic protein （GFAP） were detected by immunohistochemistry. The relative expression levels of epidermal growth factor receptor（EGFR）, vascular endothelial growth factor （VEGF） and matrix metalloproteinase-9 （MMP-9） proteins in intracerebral tumor tissues and normal tissues were examined by Western blotting. Results： Of twenty rats, three rats displayed left hemiplegia which was continuously getting worse, and the life spans of the three rats were 129, 174 and 187 d, respectively. The result of TRAP-PCR-ELISA showed that the telomerase activity of intracerebral tumor tissues was higher than that of intracerebral normal tissues. The expressions of PCNA and nestin proteins in intracerebral tumor tissues were both higher but the expression of GFAP was lower than those of intracerebral normal tissues. The results of Western blotting showed that among three rats with intracerebral normal tissues, the expression level of EGFR protein was increased in one rat while the expression level of VEGF protein was increased in two rats. There was no significant difference in the expression of MMP-9 protein among the three rats. Conclusion： The telomerase activity and the ability of cell proliferation of BMSCs can be enhanced and the activation of oncogenes can be observed after lon

关 键 词：脑肿瘤 干细胞 细胞移植 致癌性试验 

分 类 号：R379.41[医药卫生—病原生物学]