人乳腺癌细胞及其耐药细胞转录因子表达的差异

The transcription differences of mammary breast cancer cell and its adriamycin-resistant cells

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 fromMCF7/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 ceils 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 MCFT/ ADM cells is probablely due to ADM＇ s avoiding in entering the nuclei, so that ADM cannot exert function.