Ardekani Ali M, Fard Shahrzad Soleymani, Jeddi-Tehrani Mahmood, Ghahremanzade Ramin
Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran.
Avicenna J Med Biotechnol. 2011 Oct;3(4):177-93.
In many acute leukemias, normal differentiation does not occur. However, in many cell lines derived from hematologic malignancies, differentiation or apoptosis can be induced by variety of agents. Despite advances in the treatment of Acute Lymphoblastic Leukemia (ALL), in most patients long-term survival rates remain unsatisfactory, especially in T-cell derived ALL. Thus we studied the anti-cancer effects of fenretinide, 1α,25(OH)(2)D(3), and bryostatin-1 in CCRF-CEM (T-cell derived) and Nalm-6 (B-cell derived) ALL cell lines. Using MTT assays, both cell lines were shown to exhibit increased inhibition of proliferation at micro (fenretinide) and nanomolar (1α,25(OH)(2)D(3), bryostatin-1) concentrations. These anti-cancer agents were shown to induce apoptosis and activate caspase-3 pathway in both ALL cell lines. Furthermore, for the first time we are reporting consistent anti-proliferative and apoptotic effects of Bryostatin-1 in ALL T-cell derived cell line with the lowest ED(50) (ranging 4.6-7.4 nM). To evaluate the differentiation induction by fenretinide, 1α,25(OH)(2)D(3), and bryostatin-1 in ALL cell lines, we assayed for the expressions of CD19, CD38 markers on Nalm-6 and CD7 marker on CCRF-CEM cell line. The flow cytometric analysis showed a significant increase in expression of CD markers in response to anti-cancer drug treatments. To assay the effects of anti-cancer drugs on cell cycle distribution, cell cycle analysis using flow cytometry was employed. These anti-cancer drugs appear to affect the CCRF-CEM and Nalm-6 cell cycles differently (G0/G1 and G2/M arrest, respectively). Overall results demonstrate that the anti-cancer agents used in this study are strong inhibitors of ALL cell proliferation and inducers of apoptosis and differentiation in vitro. These findings may be quite helpful if these drugs are to be used for differentiation therapy of ALL patients in clinics in the future. Further studies are warranted to establish the in vivo effect of these drugs particularly in patients with T-cell derived ALL.
在许多急性白血病中，正常分化并未发生。然而，在许多源自血液系统恶性肿瘤的细胞系中，多种药物可诱导分化或凋亡。尽管急性淋巴细胞白血病（ALL）的治疗取得了进展，但大多数患者的长期生存率仍不尽人意，尤其是T细胞来源的ALL。因此，我们研究了芬维A胺、1α,25(OH)₂D₃和苔藓抑素-1对CCRF-CEM（T细胞来源）和Nalm-6（B细胞来源）ALL细胞系的抗癌作用。使用MTT试验，在微摩尔浓度（芬维A胺）和纳摩尔浓度（1α,25(OH)₂D₃、苔藓抑素-1）下，两种细胞系均显示出增殖抑制作用增强。这些抗癌药物在两种ALL细胞系中均显示出诱导凋亡并激活caspase-3途径。此外，我们首次报道了苔藓抑素-1在ALL T细胞来源的细胞系中具有一致的抗增殖和凋亡作用，其最低半数有效浓度（ED₅₀）为4.6 - 7.4 nM。为了评估芬维A胺、1α,25(OH)₂D₃和苔藓抑素-1对ALL细胞系的分化诱导作用，我们检测了Nalm-6细胞系上CD19、CD38标志物以及CCRF-CEM细胞系上CD7标志物的表达。流式细胞术分析显示，抗癌药物处理后CD标志物的表达显著增加。为了检测抗癌药物对细胞周期分布的影响，采用流式细胞术进行细胞周期分析。这些抗癌药物似乎对CCRF-CEM和Nalm-6细胞周期的影响不同（分别导致G0/G1期和G2/M期阻滞）。总体结果表明，本研究中使用的抗癌药物是ALL细胞增殖的强效抑制剂，并且在体外是凋亡和分化的诱导剂。如果这些药物未来用于ALL患者的分化治疗，这些发现可能会非常有帮助。有必要进一步研究以确定这些药物的体内作用，特别是在T细胞来源的ALL患者中。
