TNF-α增强多药耐药基因1 对骨髓保护的研究*

刘 伟; 罗 庆; 金先庆

doi:10.3969/j.issn.1000-8179.2010.03.005

TNF-α增强多药耐药基因1 对骨髓保护的研究*

Enhancive Effect of TNF-α on Transfection Efficiency of Ad-mediated Multidrug Resistance Gene 1 (MDR 1) in Mononuclear Cells of Mouse

摘要

摘要: 目的：探讨通过采用肿瘤坏死因子-α（Tumor Necrosis Factor-alpha ，TNF-α）预处理靶细胞，增强腺病毒转染小鼠单个核细胞的效率，从而增强多药耐药基因保护骨髓的能力，实现化疗中对骨髓的保护。方法：采用AdEasy-1 腺病毒载体系统重组表达MDR1 的重组腺病毒Ad-MDR 1，通过Ad5-MDR 1 感染不同浓度TNF-α 预处理后的小鼠单个核细胞，荧光倒置显微镜结合流式细胞仪监测转染率，RT-PCR 及Western blot检测TNF-α 干预后MDR1基因在靶细胞中的表达。结果：最适浓度TNF-α 预处理后，流式细胞仪检测TNF-α 预处理组腺病毒转染率（26.26%）明显高于未处理组（11.96%）；RT-PCR 和Western blot检测TNF-α 预处理组MDR1 mRNA 和P-糖蛋白（P-gp）的表达均明显高于未处理组。结论：适合浓度的TNF-α 可以明显增强腺病毒对小鼠单个核细胞的转染率，增强多药耐药基因在靶细胞中的表达，实现化疗中对骨髓的有效保护。

Abstract: Objective: To investigate the enhancive effect of TNF-α on transfection efficiency of adenovi -rus in mononuclear cells of mouse and the enhancive capability of protection of bone marrow by MDR1. Methods: Before the mononuclear cells of mouse were infected by recombinant adenovirus encoding human MDR1 gene, they had been pretreated by TNF-α. Transfection efficiency of adenovirus was moni tored by fluo-rescence microscopy, immunohistochemistry and flow cytometry (FCM). mRNA and protein levels of MDR1 in the mononuclear cells of mouse were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot before and after treatment of TNF-α. Results: After treatment of TNF-α, tansfection rates of adenovirus were obviously increased in the treated group ( 26.26% ) compared with the untreated group (11.96%). mRNA levels and protein levels of MDR1 were obviously increased in the treated group compared with the untreated group.Conclusion:TNF-α could enhance transfection efficiency of adenovi rus in mononu-clear cells of mouse and enhance capability of protection of bone marrow by MDR1.

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