MDR1在人脑肿瘤干细胞中的表达及意义

J4 ›› 2009, Vol. 35 ›› Issue (5): 769-773.

MDR1在人脑肿瘤干细胞中的表达及意义

曾飞跃, 王延金, 陈风华, 杨转移, 邓永文, 方加胜

中南大学湘雅医院神经外科|湖南长沙 410008

收稿日期:2009-03-27 出版日期:2009-09-28 发布日期:2009-09-28
通讯作者: 方加胜 E-mail:zengfeiyue@yahoo.com.cn
作者简介:曾飞跃（1977-）|男|湖南省石门县人|医学博士|主要从事胶质瘤方面的研究。
基金资助:
国家自然科学基金资助课题(30600636)

Expression and significance　of MDR1 in human brain |tumor　 stem cells

ZENG Fei-yue,WANG Yan-jin,CHEN Feng-hua,YANG Zhuan-yi,DENG Yong-wen |FANG Jia-sheng

Received:2009-03-27 Online:2009-09-28 Published:2009-09-28

摘要/Abstract

摘要：

目的：探讨脑肿瘤干细胞(BTSCs)上多药耐药基因1（MDR1）及其编码蛋白P-gp的表达，为抗癌治疗寻找靶向分子。方法：取10例胶质母细胞瘤患者肿瘤组织标本，制成单细胞溶液，接种于含EGF、bFGF、B27的无血清培养基中培养。分离获取BTSCs，对其传代培养，期间用Dispase消化分离获取到肿瘤干单细胞。免疫荧光方法检测BTSCs特异性标志物CD133，并用免疫组织化学和荧光的方法检测MDR1的表达。Western-blotting检测MDR1编码蛋白P-gp的表达。结果：所有的标本均培养出BTSCs，可在体外增殖和扩增分化，表达特异性的标志物CD133，通过免疫组织化学和免疫荧光结果提示，BTSCs上存在MDR1，MDR1在BTSCs的表达率为(45.3±5.4)%， Western blotting结果表明BTSCs上表达P-gp。结论： BTSCs表达MDR1，并且其上表达P-gp，这可能是使BTSCs更耐受化疗的原因之一。

关键词: 肿瘤干细胞；脑；多药耐药基因；P-糖蛋白

Abstract:

Abstract:Objective To investigate the expressions of MDR1 and P-glucose protein (P-gp) in brain tumor stem cells(BTSCs),and find new therapeutic targets and better anticancer strategies.Methods Tumor samples were obtained from consenting patients,actutely dissociated,triturated into single cells,and then inoculated into serum-free medium.After the primary brain tumor spheres generated,they were triturated again and passaged into fresh medium.Finally,they were digested into single cells with dispase.The specific marker CD133 was detected with immunofluorescent method.The expression of MDR1 in BTSCs was determined with immunohistochemical method,and the expression of P-gp was detected with Western blotting.Results The BTSCs isolated from glioblastoma,had the ability to propagate and differationate in vitro.The expression of CD133 was proved by immunocyt ochemistry and immunofluorescent method,the same with the MDR1,and the expression rate of MDR1 in the BTSCs was (45.3±5.4)%.P-gp expressed in BTSCs detected by Western blotting.Conclusion There are surely MDR1 and P-gp expressions in BTSCs,which may make them more resistant to chemical therapy.

Key words: neoplastic stem cells;brain;multi-drug resistance gene;P-glycoprotein

中图分类号:

R739.4

曾飞跃, 王延金, 陈风华, 杨转移, 邓永文, 方加胜. MDR1在人脑肿瘤干细胞中的表达及意义[J]. J4, 2009, 35(5): 769-773.

ZENG Fei-Yue, WANG Yan-Jin, CHEN Feng-Hua, YANG Zhuan-Yi, DENG Yong-Wen, FANG Jia-Sheng. Expression and significance　of MDR1 in human brain |tumor　 stem cells[J]. J4, 2009, 35(5): 769-773.