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Ad-Easy-1腺病毒载体系统重组表达MDR1的重组腺病毒Ad-MDR1,通过Ad5-MDR1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。
Objective： To investigate the enhancive effect of TNF-α on transfection efficiency of adenovirus in mononuclear cells of mouse and the enhancive capability of protection of bone marrow by MDRI. 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 monitored by fluorescence 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 adenovirus in mononuclear cells of mouse and enhance capability of protection of bone marrow by MDR1.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2010年第3期138-141,共4页
Chinese Journal of Clinical Oncology
基金
国家自然科学基金资助(编号:30330590)~~
