小鼠多药耐药白血病细胞系的建立及其生物学特性观察  

ESTABLISHMENT AND CHARACTERISTICS OF MURINE MULTI-DRUG RESISTANT LEUKEMIA CELL LINE

作　　者：费新红[1] 许小平[1] 陈莉[1] 高磊[1] 吕书晴[1] 陈少谊[1] 许晓巍[1] 

机构地区：[1]第二军医大学附属长海医院血液科,上海200433

出　　处：《白血病．淋巴瘤》2004年第2期75-78,共4页Journal of Leukemia & Lymphoma

摘　　要：目的:建立小鼠多药耐药白血病细胞系P388/VCR并研究其生物学特性。方法:小鼠淋巴细胞白血病细胞系P388,通过长期、单一、逐步提高浓度的方法建立多药耐药的白血病细胞系P388/VCR。采用悬浮细胞培养和半固体培养观察该细胞系的生长规律;流式细胞术分析细胞周期分布及柔红霉素在细胞内的积聚量;RT蛳PCR方法检测mdrla基因表达;四氮甲基唑蓝(MTT)法研究P388/VCR细胞系的耐药谱。结果:经过6个月单一、小剂量及间歇给药的方法诱导P388细胞成为多药耐药细胞系P388/VCR。与亲本细胞系P388比较,细胞生长曲线及倍增时间无明显差异,软琼脂培养集落形成率及大集落形成率差异不显著,细胞周期分析P388/VCR细胞S期比例降低。P388/VCR细胞除对原诱导药物具有耐药性外,对其他多种抗肿瘤药物如柔红霉素、高三尖杉酯碱、米托蒽醌、足叶乙苷等也具有交叉耐药性。流式细胞仪检测细胞内柔红霉素含量P388/VCR细胞平均荧光值为3.72,明显低于P388细胞的17.31。RT蛳PCR检测mdrla基因的表达结果为:P388细胞阴性,P388/VCR细胞阳性。结论:多药耐药细胞系P388/VCR具有mdrla基因表达阳性及对多种抗肿瘤药物耐药的特征。Objective: To establish a multi- drug resistant(MDR) cell line P388/VCR derived from murine lymphocyte leukemia cell line P388,compare the biological characteristics between P388/VCR and P388 cell line. Methods: P388 cells were induced with a long- term,intermittent,singular,unefficient vincristin(VCR)addition into MDR cell P388/VCR. The biological characteristice of P388/VCR were compared with P388,in respect of cell growth curve,colony formation percentage in soft agar and cell suspension,cell growth cycle and the intracellular accumulation of daunorubicin(DNR)by flow cytometer(FCM),and the expression of mdrla gene by the way of RT- PCR. The spectrum of muti- drug resistance of P388/VCR was evaluated by MTT assay. Results: MDR cell line P388/VCR was established by a long- term,intermittent,singular,unefficient VCR was added step by step into the culture medium of P388 cells. There was no significant difference of cell growth curve and colony formation percentages in soft agar between the two cell lines. The cell cycle analysis by FCM showed the percentage of P388/VCR in S phase was lower than that of P388. P388/VCR cells were found to be cross- resistant to DNR,homoharringtonine(HHT),epirubicin(EPI) and etoposide(VP16). Intracellular accumulation of DNR was significantly less than that in the parent cells. The expression of mdrla gene in P388 was negative through the detection of RT- PCR,but it was positive in P388/VCR. Conclusion: Comparing P388 to P388/VCR,the latter expressed mdrla gene and showed MDR characteristics of leukemia cell line.

关 键 词：白血病细胞 小鼠 多药耐药 长春新碱 

分 类 号：R733.7[医药卫生—肿瘤]