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目的建立顺铂诱导的人成骨肉瘤多药耐药细胞系(MG63/R)并观察其生物学特性。方法以顺铂为诱导剂,采用大剂量冲击与逐步增加剂量相结合的方法诱导人成骨肉瘤MG63细胞,建立多药耐药系MG63/R。MTT法检测药物敏感性;光镜、透射电镜观察MG63、MG63/R细胞形态及超微结构变化;生长曲线、克隆形成试验检测细胞增殖能力;流式细胞术检测细胞周期、凋亡指数、P-pg、bcl-2、P53蛋白表达。结果经顺铂186d的诱导,建立了MG63/R,其对顺铂的耐药指数为83.557±4.841,对阿霉素、长春新碱、氨甲基蝶呤、环磷酰氨亦产生不同程度的交叉耐药;光镜观察可见MG63/R细胞排列不规则,形态呈三角形、多角形及多核现象;透射电镜显示MG63/R细胞表面突起增加,粗面内质网丰富;细胞周期分析显示细胞增殖能力明显增加而细胞凋亡明显降低;MG63/R细胞P-pg、bcl-2阳性表达较MG63细胞明显增加,P53表达则明显降低。结论本研究建立了稳定的人成骨肉瘤多药耐药细胞系MG63/R,为进一步研究顺铂的多药耐药机制和耐药治疗提供了一种新的实验模型。
Objective To establish a cisplatin-induced human osteosarcoma multidrug resistance cell line (MG63 / R) and observe its biological characteristics. METHODS: Human osteosarcoma MG63 cells were induced by using cisplatin as an inducer and the combination of high dose and stepwise increase of dose to establish a multidrug resistance MG63 / R cell line. The morphological and ultrastructural changes of MG63 and MG63 / R cells were observed under light microscope and transmission electron microscope. The growth curve and clonogenic assay were used to detect the proliferation of cells. The cell cycle, apoptosis index and P -pg, bcl-2, P53 protein expression. Results Induction of cisplatin 186d induced MG63 / R and its resistance index to cisplatin was 83.557 ± 4.841. Adriamycin, vincristine, methotrexate and cyclophosphamide also produced different degrees The results of light microscopy showed that MG63 / R cells were irregularly arranged with triangular, polygonal and multinuclear phenomena. Transmission electron microscopy showed that the surface of MG63 / R cells was increased and the rough endoplasmic reticulum was abundant. Cell cycle analysis showed that cells Proliferative ability was significantly increased while apoptosis was significantly decreased. The expression of P-pg and bcl-2 in MG63 / R cells was significantly higher than that in MG63 cells, while P53 expression was significantly decreased. Conclusion This study established a stable human osteosarcoma multidrug resistance cell line MG63 / R, which provided a new experimental model for further study on the multidrug resistance mechanism and drug resistance of cisplatin.