目的: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(Saos-2/ADM1,Saos-2/ADM 4)的生物学特性.方法:以ADM为诱导剂,采用逐步增加药物剂量冲击诱导方法,诱导人成骨肉瘤原代Saos-2细胞株,建立不同药物浓度耐药系(Saos-2/ADM1,Saos-2/ADM4);MTT法检测原代与耐药细胞株对ADM、顺铂(Cispla-tin,DDP)、甲氨蝶呤(methotrexate,MTX)、异环磷酰胺(Ifos-famide,IFO)、表柔比星(Epirubicine,EPI)、比柔比星(Pirambi-cin,THP)、紫杉醇(Paclitaxel,PTX)药物敏感性;光学显微镜、透射电镜观察细胞形态及超微结构变化;细胞生长曲线检测细胞增殖能力;流式细胞仪检测细胞周期比例.结果:经167d的诱导,建立了Saos-2/ADM1,Saos-2/ADM4细胞株,其对ADM的耐药指数(Resistant drug index,RI)分别为原代细胞株49.8和74.6倍;耐药细胞株对MTX,EPI,THP,PTX亦产生不同程度的交叉耐药,对DDP仍然敏感;光镜观察Saos-2/ADM1,Saos-2/ADM4细胞株细胞体积增大,多核现象较原代Saos-2细胞株明显增加;透射电镜显示Saos-2/ADMI,Saos-2/ADM4细胞株表面突起较原代Saos-2细胞株减少、且核仁增大增多;细胞生长曲线显示耐药细胞株增殖能力下降,S期细胞所占比例减少.结论:建立了耐ADM人骨肉细胞株Saos-2/ADM1,Saos-2/ADM4并观察了耐药细胞株的生物学特性.
Establishment of adriamycin resistance cell lines of human osteosarcoma and their biological characteristics
AIM: To establish the resistant human osteosarcoma cell lines (Saos-2/ADM1 and Saos-2/ ADM4 ) from the Saos-2 cell line and test their biological characteristics. METHODS: Saos-2 cells were exposed to gradually increased doses of ADM culture medium to establish resistant cell lines ( Saos-2/ADM1 and Saos-2/ ADM4). The sensitivity of the Saos-2, Saos-2/ ADM1 and Saos-2/ ADM4 cell lines to ADM, DDP, EPI, MTX, IFO, EPI, THP and PIX was measured by MTT assay. The morphology and ultrastructure of the cell lines were observed by optical microscopy and transmission electron microscopy. The proliferation ability of the cells was measured by growth curve. The cell cycles were analyzed by flow cytometry. RESULTS: Resistant cell lines (Saos-2/ADM1 and Saos-2/ ADM4) were established after induction for 167 days. The resistance index of the Saos-2/ ADM1 and Saos-2/ ADM4 cell lines to ADM was 49. 8 and 74.6 times more than Saos-2 respectively. The two cells lines had resistance to MTX, EPI, THP and PTX. The cells remained sensitive to DDP. Disordered structure of the Saos-2/ADM1 and Saos-2/ ADM4 cells was observed through microscopy. The cells appeared coenocytically. The decrease of cell villus and the increase of nucleoli were observed through transmission electron microscopy. The proliferation ability of Saos-2/ADM 1, Saos-2/ ADM4 cells decreased significantly. The cells in S stage decreased. CONCLUSION: Resistant adriamycin cell lines (Saos-2/ADM1 and Saos-2/ ADM4 ) of human osteosarcoma are established and their biological characteristics are investigated.
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万方数据
维普
