目的 研究阿霉素在不同药敏性肿瘤细胞中转录因子水平的差异,为肿瘤细胞多药耐药机制的研究提供参考.方法 通过细胞活力检测野生型乳腺癌细胞(MCF7/W)和对阿霉素耐药的乳腺癌细胞(MCF7/ADM)的药敏性差异;荧光显微镜镜检阿霉素荧光分布;有机溶剂萃取法进行全细胞阿霉素吸收分析;细胞膜和核膜分离检测阿霉素的亚细胞定位;通过转录因子膜分析MCF7/W和MCF7/ADM细胞中转录因子表达差异.结果 两株细胞的耐药IC50相差近10 000倍左右;利用镜检和定量分析,发现阿霉素在MCF7/W细胞中主要定位于胞核,而在MCF7/ADM细胞中则蓄积较少且主要定位于胞质;转录因子差异分析发现膜上总345个转录因子中,与野生型细胞相比,耐药细胞的106个转录因子表达上调,32个表达下调.结论 阿霉素在耐药细胞与野生型细胞中转录因子表达的差异,可能是干扰阿霉素入核,阻止其功能发挥的基础.%Aim To investigate the transcription difference of adiramycin( ADM ) in cells with different drug sensitivity, and from which to provide the evidence for multidrug resistance. Methods The different drug sensitivity of MCF7/W and MCF7/ADM cells was measured by cell vitality assay; fluroescence microscopy was used to detect the different localization of the two cells which were treated with the same dose of ADM; ADM was extract by organic solvent fromrnMCF7/W and MCF7/ADM respectively and the content was assayed;the cell membrane and nuclear membrane were separated for assaying the localization of ADM. From the protein/DNA array, the transcription differences of the two cells were discovered. Results IC50 of MCF7/ADM over IC50 of MCF7/ADM was nearly 10 000; microscope detection and quantitative analysis displayed that the localization of ADM in MCF7/W was mainly in the nuclei , but in MCF 7 / ADM cells , ADM accumulation was much lower and mainly localized in the cytoplasm. From protein/DNA array, 106 transcription factors among the total 345 transcription factors were up-regulated, while 32 transcription factors were down-regulated. Conclusions The difference of transcription factors in MCF7/W and MCF7/rnADM cells is probablely due to ADM' s avoiding in entering the nuclei, so that ADM cannot exert function.
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