Robinson Blaine W, Behling Kathryn C, Gupta Manish, Zhang Alena Y, Moore Jonni S, Bantly Andrew D, Willman Cheryl L, Carroll Andrew J, Adamson Peter C, Barrett Jeffrey S, Felix Carolyn A
Division of Oncology, The Children's Hospital of Philadelphia, Philadelphia, PA, USA.
Br J Haematol. 2008 Jun;141(6):827-39. doi: 10.1111/j.1365-2141.2008.07100.x. Epub 2008 Apr 18.
Chemotherapy resistance from imbalanced apoptosis regulation may contribute to poor outcome in leukaemias with t(4;11). Anti-apoptotic BCL-2 expression and target modulation were characterized in cell lines with t(4;11) and BCL-2 expression was examined in MLL and non-MLL infant/paediatric leukaemia cases by Western blot analysis and/or real-time polymerase chain reaction. Cytotoxicity of Genasensetrade mark (Oblimersen Sodium, G3139) alone or combined with cytotoxic drugs was assessed by MTT [(3-4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] assays of the cell lines, applying pharmacostatistical response surface modelling of drug interactions. Apoptosis and cell cycle were evaluated by flow cytometry in RS4:11 cells. Primary leukaemias and cell lines with t(4;11) expressed abundant BCL2 mRNA and protein. Variable, sometimes substantial BCL2 mRNA was detected in other leukaemia subtypes. G3139 reduced BCL2 mRNA and protein in RS4:11 cells. The most sensitive cell line to single-agent G3139 was RS4:11. Low G3139 concentrations sensitized RS4:11 and MV4-11 cells to select anti-leukaemia cytotoxic drugs. In RS4:11 cells, combining G3139 with doxorubicin (ADR) increased active caspase 3 and TUNEL staining compared to ADR alone, indicating greater apoptosis, and G3139 increased S-phase progression. The abundant BCL-2 affords a molecular target in leukaemias with t(4;11). G3139 exhibits preclinical activity and synergy with select cytotoxic agents in RS4:11 and MV4-11 cells, and these effects occur through apoptosis.
凋亡调节失衡导致的化疗耐药可能是t(4;11)白血病预后不良的原因。对具有t(4;11)的细胞系进行抗凋亡BCL-2表达及靶点调控特征分析,并通过蛋白质免疫印迹分析和/或实时聚合酶链反应检测MLL及非MLL婴儿/儿童白血病病例中的BCL-2表达。通过MTT [(3-4,5-二甲基噻唑-2-基)-2,5-二苯基溴化四氮唑]法检测细胞系中Genasensetrade mark(奥布利森钠,G3139)单独或与细胞毒性药物联合使用时的细胞毒性,并应用药物相互作用的药效统计学响应面模型。通过流式细胞术评估RS4:11细胞中的凋亡和细胞周期。原发性白血病和具有t(4;11)的细胞系表达丰富的BCL2 mRNA和蛋白。在其他白血病亚型中检测到可变的、有时大量的BCL2 mRNA。G3139可降低RS4:11细胞中的BCL2 mRNA和蛋白水平。对单药G3139最敏感的细胞系是RS4:11。低浓度G3139可使RS4:11和MV4-11细胞对选定的抗白血病细胞毒性药物敏感。在RS4:11细胞中,与单独使用阿霉素(ADR)相比,G3139与阿霉素联合使用可增加活化的半胱天冬酶3和TUNEL染色,表明凋亡增加,且G3139可增加S期进程。丰富的BCL-2为t(4;11)白血病提供了一个分子靶点。G3139在RS4:11和MV4-11细胞中表现出临床前活性并与选定的细胞毒性药物具有协同作用,且这些作用通过凋亡发生。