载脂蛋白A-Ⅱ与阿霉素耐药的相关性研究

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目的 检测载脂蛋白A-Ⅱ(apolipoprotein A-Ⅱ,APOA-Ⅱ)与阿霉素(adriamycin,ADM)耐药的相关性.方法 IC50分析细胞对 ADM 的敏感性;激光共聚焦检测 MCF7/W 细胞、MCF7/ADM细胞及APOA-Ⅱ高表达的MCF7/W细胞内ADM的分布;RT-PCR和Western blot分别检测MCF7/W和MCF7/ADM细胞内APOA-Ⅱ基因及蛋白的表达;IC50分析高表达APOA-Ⅱ的HEK293细胞的药敏性以及高表达APOA-Ⅱ的乳腺癌细胞 T47D、MDA-MB231的药敏性.结果MCF7/ADM细胞与MCF7/W细胞相比,耐药指数达13.0(P<0.05);MCF7/W细胞中的ADM在细胞核中大量聚集,而MCF7/ADM细胞中ADM则几无细胞核分布;同时,高表达APOA-Ⅱ蛋白的MCF7/W细胞,其细胞核中ADM的分布明显少于正常表达APOA-Ⅱ的MCF7/W细胞;MCF7/ADM细胞中APOA-Ⅱ的基因及蛋白表达水平均高于亲本细胞(P<0.05);转染了APOA-Ⅱ的HEK293细胞对ADM的敏感性明显降低(P <0.05);并且转染了 APOA-Ⅱ的乳腺癌细胞T47D、MDA-MB231对ADM的药敏性也降低(P<0.05).结论 APOA-Ⅱ蛋白与乳腺癌ADM耐药相关,为临床ADM的化疗提供了一定的指导意义.“,”Aim To study the relationship between apolipoprotein A-Ⅱ (APOA-Ⅱ) and adriamycin re-sistance. Methods The drug sensitivity of cells was analyzed by IC50assay. The localizations of ADM in MCF7/W,MCF7/ADM and APOA-Ⅱ transgenic cells were observed by laser scanning confocal microscopy. RT-PCR and Western blot were applied to detect the gene and protein expression in MCF7/W and MCF7/ADM cells, respectively. The APOA-Ⅱ transgenic HEK293 cells were analyzed by IC50assay,the IC50of breat cancer cells T470 and MDA-MB231 were ana-lysed as well. Results Compared with MCF7/W cells,the resistance index of MCF7/ADM cells got 13. 0 (P<0.05). ADM localized mainly in the nuclei of MCF7/W cells,but none in the nuclei of MCF7/ADM cells. Meanwhile,the localization of ADM in APOA-Ⅱtransgenic MCF7/W cells was obviously less than that in normal parent MCF7/W cells. The gene and protein expression levels of APOA-Ⅱ in MCF7/ADM cells were both higher than in MCF7/W cells (P<0.05);the sensitivity of APOA-Ⅱtransgenic HEK293 to ADM was also reduced (P<0.05). Furthermore,the sensi-tivity of breast cancer cells T47D and MDA-MB231 to ADM was reduced as well (P<0.05). Conclusion APOA-Ⅱ is found to be positively related to ADM re-sistance,which would provide instruction for the clini-cal usage of ADM.
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