中文摘要：

目的：建立人弥漫大B细胞淋巴瘤（DLBCL）阿霉素（ADM）耐药细胞株，为研究DLBCL耐药机制提供模型。方法：以体外低浓度梯度递增法诱导筛选人DLBCL耐ADM细胞株Pfeiffer／ADM。在显微镜下观察细胞形态变化，绘制生长曲线，计算倍增时间。CCK-8法检测Pfeiffer／ADM对不同药物的耐药系数（RI）及脱药1个月后的耐药稳定性。在含补体的血清中，CCK-8法比较两细胞株对利妥昔单抗（RTX）介导的补体依赖性细胞毒性（CDC）的差别，荧光定量PCR（qRT-PCR）法检测两株细胞中多药耐药基因1（MDR1）tuRNA相对表达量。结果：经过约7个月的诱导，成功构建耐ADM细胞株Pfeiffer／ADM；显微镜下Pfeiffer和Pfeiffer／ADM细胞形态无明显差别，亲代加药后死亡细胞更多，而耐药株生长依旧良好；Pfeiffer和Pfeiffer／ADM的倍增时间分别为（32．28±1．18）h和（31．41±2．03）h，两者比较差异无统计学意义（P〉0．05）；通过CCK-8法测得Pfeiffer／ADM对ADM、依托泊苷（VP-16）和长春新碱（VCR）的耐药指数（RI）分别为12．87、13．18、16．95；脱药1个月和冻存复苏后，Pfeiffer／ADM依旧能保持耐药性，其对ADM的Ic。。值分别为0．52mg／L、0．49mg／L；两细胞株对于不同浓度的RTX介导的CDC效应无明显差别（P〉o．05）；Pfeiffer／ADM细胞的MDR1mRNA相对表达量明显高于Pfeiffer细胞株（P〈0．05）。结论：成功建立具有多药耐药性的DLBCL Pfeiffer／ADM细胞株，化疗耐药的Pfeiffer／ADM细胞与RTX无交叉耐药性，该细胞是研究DLBCL获得性耐药的机制和逆转耐药的理想模型。

英文摘要：

Objective： To establish adriamycin-resistant human diffuse large B-cell lymphoma （DLBCL） cell lines. Methods： Human DLBCL cells （Pfeiffer） were exposed in stepwise escalating concentration of adria- mycin until the resistant cell line （Pfeiffer/ADM） was developed. The morphology and cell doubling time of Pfeiffer and Pfeiffer/ADM cells were compared under the microscope. The drug resistance fold, stability and rituximab （RTX） -mediated complement-dependent cytotoxicity were detected by CCK-8 assay. The multidrug resistance 1 （MDR1） mRNA expression was detected by quantitative real-time PCR （qRT- PCR）. Results： Pfeiffer/ADM cell line was established after 7 months of induction. In a medium contained adriamycin, Pfeiffer/ADM grew well, but most of Pfeiffer cells were dead. The doubling time of Pfeiffer and Pfeiffer/ADM cells were （32.28±1.18） h and （31.41±2.03） h, respectively （ P〉0.05）. CCK-8 as- say showed that the indexes of drug resistant to adriamycin, etoposide and vincristine in Pfeiffer/ADM were 12.87, 13.18 and 16.85, respectively. Pfeiffer/ADM remained drugs resistant in drug-free medium atleast 1 month or after the recovery from the frozen. There was no significant difference between Pfeiffer and Pfeiffer/ADM in RTX-mediated complement- dependent cytotoxicity （ P〉0. 05）. Pfeiffer/ADM was significantly higher than Pfeiffer in MDR1 mR-NA expression （ P〈0.05）. Conclusion： A DLBCL multi-drugs resistant fully, which was not cross-resistance to RTX. It could be an ideal model cell line was established success- to study mechanisms of acquired drug resistance and reverse drug resistance.