【作者】 张莲芬； 严婷； 王妍； 冯磊； 金坚；

【Author】 Lian-Fen Zhang,Ting Yan, Yan Wang, Lei Feng, Jian Jin* (Department of Pharmaceupical and Molecular Biotechnology, School of Biotechnology, Southern Yangtze University, Wuxi, PR China 214036 )

【机构】 江南大学生物工程学院生物制药系；

【摘要】 采用低浓度加量持续诱导法建立了耐阿霉素(ADM)的人乳腺癌细胞系MCF-7/ADMy,并对其生物学特性和阿霉素结合蛋白进行了初步分析评价。结果表明,MCF-7/ADMy细胞较亲本细胞的ADM的IC50高500倍,撤药培养150天后耐药指数仍维持在200倍以上,并对多种化疗药物产生耐药性;耐药细胞分化程度低于同步传代的MCF-7细胞,细胞倍增时间与亲本细胞接近,S期细胞显著增加,G1期细胞减少;随撤药时间的延长,细胞的增殖速度加快;耐药细胞P-gP、LRP和GSTπ的表达水平较亲本细胞有显著增加,ER阳性表达丢失;在稳定生长的撤药培养6天的细胞中仍有ADM蓄积。亲和甄别磁珠法从MCF-7和MCF-7/ADMy细胞全蛋白中分离出与ADM结合的蛋白,两株细胞中均有多种与ADM结合的蛋白,其中主要的结合蛋白相同,均为actin,myosin和keratin。本实验建立的MCF-7/ADMy模型具有多药耐药细胞的基本生物学特性,ADM结合的蛋白分析提示其还可能作用于细胞的骨架结构蛋白来干扰细胞的有丝分裂过程,达到抑制肿瘤细胞增殖的目的。更多还原

【Abstract】 In this paper a line of breast cancer cell named MCF-7/ADMy with multidrug-resistance was established, and its elementary biological properties including drug-resistance, cell cycle dynamics, exterior transformation, intracellular drug accumulation, the binding protein with ADM and so on were analyzed. Results showed that MCF-7/ADMy was of good resistance to several chemotherapeutants for tumor. The IC50 of MCF-7/ADMy to ADM was 500 times higher than that of nature MCF-7, and its IC50 to other chemotherapyagents was 6-75 times higher too. However, the drug resistant index of the cells could keep on over 200 times higher after culturing MCF-7/ADMy in the medium (without ADM) for 150 days. The multiplication time of MCF-7/ADMy was similar to that of their nature cells, but the most cells concentrated in S-phase of the cell cycle and the least cells in G1-phase. The multiplication time could be accelerated in evidence with culturing the cells in the medium (without ADM). The expression levels of P-gp、LRP and GSTπ on the cells increased significantly except for ER. The level of intracellular ADM concentration of the cells remained high quantity even after 6-day culture in the medium (without ADM). The protein which could bind with ADM on the two cell lines was isolated by the ADM affinity screening. The major binding protein were the same on the two cell lines,such as the actin, the myosin, and the keratin. The results indicated that the multidrug-resistant cell species (MCF-7/ADMy) established in this study was of good biological characteristics, multidrug-resistance especially, which would be helpful to research on MDR mechanism of cancer cells. The mechanism of ADM to inhibit the proliferation of tumor cell may be caused by the interventing the mitosis through interaction with the cystoskeleton.更多还原