脑胶质瘤细胞体外促进人骨髓间充质干细胞肿瘤相关基因表达  

Glioma cells promote expression of cancer-related genes in human bone marrow-derived mesenchymal stromal cells in vitro

作　　者：朱汝森[1] 许成杰[1] 兰柳波[2] 陈兴贵[3] 梁远生[1] 尹延庆[1] 

机构地区：[1]广东医学院附属医院神经外科,湛江524001 [2]广东医学院生物化学与分子生物学研究所 [3]广东医学院附属医院肿瘤中心

出　　处：《中国神经精神疾病杂志》2016年第1期50-55,共6页Chinese Journal of Nervous and Mental Diseases

基　　金：广东省自然科学基金(编号:2015A030313525);广东省医学科研基金(编号:B2009197);湛江市科技攻关计划项目(编号:2014B01087);广东医学院博士科研基金

摘　　要：目的观察与人脑胶质瘤细胞共培养后的人骨髓间充质干细胞(human bone marrow-derived mesenchymal stromal cells,h MSCs)中肿瘤相关基因表达的变化,初步评估h MSCs在人脑胶质瘤环境中的生物安全性在致瘤性方面的风险。方法将h MSCs与人脑胶质瘤细胞U251于体外共同培养5d后,通过肿瘤基因芯片、实时定量RT-PCR及Western-blot实验检测h MSCs中肿瘤相关基因表达水平的变化。结果基因芯片结果显示,与单独培养的h MSCs对比,与人脑胶质瘤细胞U251共同培养后的h MSCs中存在SPINT2、TK1、STC1、MMP1、CCND1、SORT1、SEPT6、CDC20、SHB、CDK5、RELA、XRCC4、KIT、CTPS、CAPNS1及ETV6等16个肿瘤相关基因的表达明显上调(3倍以上),而无一基因表达下调;实时定量RT-PCR结果显示癌基因KIT、CAPNS1、TK1、MMP1、CCND1、CDC20、RELA以及STC1在共培养组h MSCs中呈表达上调;Western-blot结果显示癌基因KIT、MMP1、CCND1以及RELA的蛋白水平在共培养组h MSCs中呈表达上调。结论 h MSCs在脑胶质瘤细胞的影响下可出现部分癌基因的高度表达,提示了对于h MSCs应用于对脑胶质瘤进行基因治疗的生物安全性在致瘤性方面的风险需引起关注及深入研究。Objective We investigated the expression profile of cancer related genes in hMSCs co-cultured with U251 glioma cells, to evaluate the risk of malignant transformation of hMSCs in glioma environment. Methods hMSCs were co-cultured with U251 glioma cells for 5 days and the expression profile of cancer-related genes were investigated by using microarray assay, followed by Real-time quantitative RT-PCR and Western blot. Results Of the 440 cancer-re- lated genes covered by Oligo GEArray Human Cancer Microarray OHS-802, SPINT2, TK1, STC1, MMP1, CCND1, SORT1, SEPT6, CDC20, SHB, CDK5, RELA, XRCC4, KIT, CTPS, CAPNS1 and ETV6 were significantly upregulated （〉 3-fold） whereas none was downregulated in hMSCs co-cultured with U251 glioma cells. The upregulation of oncogenes KIT, CAPNS1, TK1, MMP1, CCND1, CDC20, RELA and STC1 in co-cultured hMSCs were confirmed by Real-time quan-titative RT-PCR. The upregulation of protein expression of oncogenes KIT, MMP1, CCND1 and RELA were detected by Western blot. Conclusion The present study demonstrates that co-culture of hMSCs with human glioma cells leads to up- regulation of some important oncogenes in hMSCs, indicating the tumorigenic potential of hMSCs in glioma environment.

关 键 词：骨髓间充质干细胞 脑胶质瘤基因治疗 致瘤性肿瘤相关基因 基因表达 

分 类 号：R739.4[医药卫生—肿瘤]