【摘要】 目的:建立对马法兰和阿霉素耐药的人骨髓瘤细胞系。方法:在人骨髓瘤细胞系RPMI8226培养体系中分别加入1.0×10-8M的阿霉素(DOX)和1.0×10-6M的马法兰(LP),每周3次换液,DOX于10月后增加至1.0×10-7M,LP于47周后增加至5×10-6M。MTT法测定敏感细胞系和耐药系8226/DOX的IC50值,在耐LP细胞系8226/LP中加入丁硫氨酸亚砜胺(BSO)阻断谷胱甘肽的合成,MTT法测定细胞系8226/S、8226/LP细胞及8226/LP+BSO细胞的IC50值。HE染色普通光学显微镜下观察各组细胞形态,流式细胞仪检测各组细胞表面P-糖蛋白、CD38、CD45、CD19。台盼蓝拒染法测出8226/S、8226/DOX、8226/LP细胞的生长曲线。结果:所培养耐药细胞系8226/DOX、R8226/LP具有亲母细胞系的生物学特点,形态相似,均为CD38+CD45-CD19-表达细胞,倍增时间与亲母细胞系相近。比较IC50,8226/DOX对DOX的耐药约15倍于亲母细胞系。8226/LP对LP的耐药约7倍于亲母细胞系,而经过BSO阻断,其敏感性明显增加。结论:建立的骨髓瘤细胞系RPMI8226/LP、RPMI8226/DOX可作为骨髓瘤耐药研究的有效模型。更多还原

【Abstract】 Objective: To develop two human multiple myeloma cell lines which are resistant to doxorubicin(DOX) and melphalan(LP) respectively. Methods: RPMI8226(8226/S) cells were intially exposed to DOX at a concentration of 1.0×10-8 M and LP at a concentration of 1.0×10-8 M respectively. The DOX concentration was gradually increased to a final concentration of 1.0×10-7 M over 10-month period. The LP concentration was gradually increased to a final concentration of 5×10-6 M over 47-week period. Determine IC50 of 8226/S and 8226/DOX by MTT. Buthionine sulfoximine(BSO) was used to deplete glutathione levels in the 8226/LP cells. A relative resistance index was expresssed as the ratio of the IC50 of the resistant cells to the IC50 of the sensitive cells (8226/S). Observing the morphology of each group cells under routine microcopy by HE stainning. Flowcytometry was used to investigate P-glycoprotein, CD38, CD45 and CD19. The growth curves were determined by trypan blue dye exclusion. Results: The drug-resistant cell lines 8226/DOX and 8226/LP we developed had the biological characteristic of parent cell line. They all were CD38+CD45-CD19- cells. Their morphology was similar. Comparing IC50, The resistance of 8226/DOX was 15 fold approximately of that of 8226/S, 8226/LP was 7 fold approximately of 8226/S. After inhibited by BSO, the sensitivity to LP of 8226/LP increased obviously. Conclusions: The human multiple myeloma cell lines 8226/DOX and 8226/LP we developed are potent tools for studies of drug-resistance of multiple myeloma.更多还原