耐阿霉素人骨肉瘤细胞株的建立及评价 - 第三军医大学学报.doc

人骨肉瘤耐阿霉素细胞株的建立 陈渝 邓忠良* （重庆医科大学附属第二医院重庆400010诱导并建立耐阿霉素(Adriamycin，ADM)的人骨肉瘤细胞株，观察耐药细胞系()的生物学特性采用逐步递增ADM浓度间歇作用的方法诱导法检测药物敏感性观察细胞形态变化；生长曲线、检测细胞；流式细胞术检测细胞周期甲氨喋啶异环磷胺长春新碱紫杉醇143B/ADM细胞细胞核增大143B/ADM克隆形成能力显著升高。结论 本研究建立了稳定的人骨肉瘤多药耐药细胞系()，为进一步研究多药耐药机制和耐药治疗提供了一种新的实验模型。；；，Establishment of adriamycin resistance cell lines of human osteosarcoma Chen Yu，（ Department of , the Second Affiliated Hospital, ChongqingMedical University, Chongqing 400010, China）To establish a multidrug resistance cell line from human osteosarcoma 143B cell line and study characterization of the cell line. Methods The 基金项目：国家自然科学基金（） Supported by the National Natural Science Foundation of China（） 作者简介：，，，主治医师，从事研究通信作者：，E-mail: deng7586@ human osteosarcoma 143B cells were exposed to high and gradually increasing dose of adriamycin for 45 d to establish muhidrug resistance cell line 143B/ADM. The sensitivity of muhidrug resistance was measured by CCK assay. The cell stability was measured by cryopreservation/resuscitation test and withdrawal test. Morphology was observed by laser scanning confocal microscope. The ability of proliferation was measured by growth curve, adherence ratio and double time. Cell cycle was analyzed by flow cytometry. Clonality was detected by soft agar colony formation test. Results Resistant cell line(143B/ADM) was established after induction for 45 d. The resistance index of 143B/ADM cells to adriamycin was 15.7, and the cells showed various resistance to cisplatin, methotrexate, isophosphamide, vincristine and taxinol. Cryopreservation/resuscitation and withdrawal had no effect on resistance. Laser scanning confocal microscope observed that the volume of cell and nuclei of 163B/ADM was increased. The proliferation ability of 143B/ADM cells decreased significantly, and the clonality of 143B/ADM cells increased significantly. Flow cytometry showed the cell cycle of 143B/ADM was disordered. Conclusion A multi drug resistanc