耐糖皮质激素人类弥漫大B细胞淋巴瘤细胞系的建立及其生物学特性观察  

Establishment and biological characterization of a human glucocorticoid-resistant cell line of diffuse large B cell lymphoma

作　　者：谭微[1] 陈波斌[1] 马燕[1] 许小平[1] 林果为[1] 

机构地区：[1]复旦大学附属华山医院血液科,上海200040

出　　处：《白血病．淋巴瘤》2013年第12期712-716,共5页Journal of Leukemia & Lymphoma

基　　金：上海市自然科学基金（10ZR1405200）

摘　　要：目的 采用长期间歇小剂量糖皮质激素（GC）递增诱导法建立耐GC的人类弥漫大B细胞淋巴瘤细胞系Toledo／地塞米松（DEX）,并观察耐药和亲本细胞株两者之间的生物学特性差异,探讨其耐药机制.方法 以DEX为诱导剂,将对数期亲本Toledo细胞接种在含DEX起始浓度为1×10-s mol／L的培养液中培养96 h后,改为不含药物的培养液培养,待细胞增殖再次进入对数生长期,提高DEX浓度1倍,直至在含DEX 1.024×10-5 mol／L的浓度中稳定生长.并通过细胞学、遗传学、免疫学、分子生物学和裸鼠致瘤实验等多种方法观察耐药和亲本细胞系两者之间的生物学特性差异,探讨GC耐药机制.结果 Toledo／DEX细胞系可以在含DEX 1.024×10-5 mol／L的浓度中稳定生长.耐药及亲本细胞系在形态及免疫表型上完全一致.对超微结构观察可见Toledo／DEX细胞表面局部可见片层状突起,有细长微绒毛.耐药细胞致瘤性强,对多种化疗药有耐药性且耐药指数高,提示耐药细胞更具侵袭性.耐药细胞增殖慢,较多被阻滞在G1期,染色体变异多,但未形成两个相关克隆.此外,GC受体（GR）α、GR β表达异常参与GC耐药的形成.结论 Toledo／DEX是一株生物学特性稳定的高耐药细胞系,为研究GC耐药机制及探索逆转GC耐药性的药物提供了生物学基础.Objective To establish a novel glucocorticoid （GC）-resistant human diffuse large B lymphoma（DLBCL） cell line Toledo/dexamethasone （DEX） by the exposure to DEX,and observe the biological characteristics of resistant and parental cell line,investigate the mechanisms of glucocorticoid-resistance.Methods Toledo/DEX was established by the exposure to DEX,the dose of which was increased gradually and intermittently for long periods of time.Toledo,in the logarithmic growth phage,was incubated in the culture medium containing DEX at the concentration of 1×10-8 mol/L at first.The medium without DEX was replaced after for 96 hours until the cell line re-entered the logarithmic growth phase.Repeat the above steps to acquire the ultimate concentration of DEX in the medium as 1.024 ×10-5 mol/L.The biological characteristics of resistant and parental cell lines were evaluated.Results Toledo/DEX was more invasive in the aspects of ultrastructure,tumorigenicity and drug sensitivity.Meanwhile,Toledo/DEX achieved some stable biological characteristics such as morphology,karyotypes and immunophenotype.Furthermore,GC receptor （GR） α and GR β protein expression analysis showed that GR was involved in the mechanism of the GCresistance.Conclusions Toledo/DEX is a drug-resistant cell line with a stable biology backgroud.These results may help shed light on the knowledge of GC-resistance and lay the groundwork for searching new therapeutics to reverse drug-resistance.

关 键 词：淋巴瘤 B细胞 细胞系 糖皮质激素 耐药 

分 类 号：R733.1[医药卫生—肿瘤]