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mdr1及P-gp与脑胶质瘤耐药关系的研究

付旭东   

  1. 郑州大学第五附属医院神经外科
  • 收稿日期:2010-05-10 修回日期:2010-09-10 出版日期:2011-02-15 发布日期:2011-02-15
  • 通讯作者: 付旭东

A Study on the relation between mdr1,P-gp and multidrug resistance of human glioma

  • Received:2010-05-10 Revised:2010-09-10 Published:2011-02-15 Online:2011-02-15

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2001

摘要： 摘要 目的 探讨脑胶质瘤细胞多药耐药(MDR)形成的机制。方法 体外培养人脑胶质瘤细胞系U251及多药耐药细胞系U251/Dox。MTT法检测U251/Dox细胞对4种化疗药物耐药倍数；流式细胞术检测U251、U251/Dox细胞对罗丹明123(Rh123)蓄积与泵出能力；分别采用RT-PCR和免疫组织化学法检测U251、U251/Dox多药耐药基因(mdr1)和蛋白的表达水平。结果 U251/Dox细胞对4种化疗药物均产生耐药性；Rh123的蓄积和泵出试验显示，U251/Dox比U251细胞的平均荧光强度(MFI)显著减少，P<0.05；其mdr1 mRNA灰度比值比U251细胞均明显增加，P<0.05；U251/Dox细胞P-gp表达显著高于U251细胞。结论 脑胶质瘤多药耐药产生机制与mdr1 mRNA过度表达和影响P-糖蛋白(P-gp)的功能有关。

关键词: 脑胶质瘤, 多药耐药, P-糖蛋白, 流式细胞术, RT-PCR

Abstract: Abstract: Objective: To study the mechanism of multidrug resistance of human glioma. Methods：The human glioma cells U251 and multidrug resistance cells U251/Dox were cultured in vitro. Accumulation and efflux ability of Rhodamine123 (Rh123) in U251 and U251/Dox cells were determined by flow cytometry；Multidrug resistance gene 1(mdr1) mRNA expression level in U251 and U251/Dox cells were measured by semi-quantitative reverse transcription and polymerase chain reaction (RT-PCR). Expression of P-gp was detected by immunohistochemistry. Results：MFI of U251 and U251/Dox cells were different；The mdr1 mRNA level in U251/Dox cells increased significantly. The expression of P-gp in U251 cells was significantly lower than U251/Dox cells. Conclusion: Human glioma develop cross-resistance to multiple tested cytotoxic agents and the possible molecular mechanism is involved with mdr1 mRNA overexpression and influence the function of P-glycoprotein.

Key words: Glioma, Multidrug resistance, P-gp, Flow cytometry, RT-PCR