Ji Jie, Valdez Benigno C, Li Yang, Liu Yan, Teo Esmeralda C, Nieto Yago, Champlin Richard E, Andersson Borje S
Department of Stem Cell Transplantation & Cellular Therapy, University of Texas M. D. Anderson Cancer Center, Houston, TX.
Department of Stem Cell Transplantation & Cellular Therapy, University of Texas M. D. Anderson Cancer Center, Houston, TX.
Exp Hematol. 2016 Jun;44(6):458-65. doi: 10.1016/j.exphem.2016.03.001. Epub 2016 Mar 11.
Hematopoietic stem cell transplantation (HSCT) is an effective treatment for patients with refractory lymphomas. Nucleoside analogs (NAs) and DNA alkylating agents are efficacious in treating hematologic malignancies. To design an efficacious and more economical pretransplant regimen for lymphoma patients, we analyzed the cytotoxicity of cladribine (Clad), gemcitabine (Gem), busulfan (Bu), and suberoylanilide hydroxamic acid (SAHA) in lymphoma cell lines. J45.01 and U937 lymphoma cell lines were exposed to drugs, alone or in combination, for 48 hours and analyzed with the MTT and annexin V assays, Western blotting, and flow cytometry. On the basis of the IC5-10 values of the drugs, the Clad+Gem+Bu combination inhibited the proliferation of both cell lines to ∼55%-60%. Addition of SAHA to this combination decreased proliferation further to ∼30%. Exposure to the Clad+Gem+Bu+SAHA combination activated the DNA damage response and ATM-CHK2 pathway; modified histones; decreased mitochondrial membrane potential, which caused leakage of apoptosis-inducing factors; and activated apoptosis. Pretreatment of cells with the pan-caspase inhibitor Z-VAD-FMK blocked the phosphorylation of histone 2AX and cleavage of PARP-1 and caspases. The Clad+Gem+Bu+SAHA combination provides synergistic cytotoxicity in lymphoma cell lines. Our results may be a basis for using this combination as a pretransplant conditioning regimen in a clinical trial for lymphoma patients undergoing hematopoietic stem cell transplantation, replacing the more expensive nucleoside analog clofarabine.
造血干细胞移植（HSCT）是治疗难治性淋巴瘤患者的一种有效方法。核苷类似物（NAs）和DNA烷化剂在治疗血液系统恶性肿瘤方面有效。为了为淋巴瘤患者设计一种有效且更经济的移植前方案，我们分析了克拉屈滨（Clad）、吉西他滨（Gem）、白消安（Bu）和辛二酰苯胺异羟肟酸（SAHA）在淋巴瘤细胞系中的细胞毒性。将J45.01和U937淋巴瘤细胞系单独或联合暴露于药物48小时，并用MTT和膜联蛋白V检测、蛋白质印迹法和流式细胞术进行分析。根据药物的IC5 - 10值，Clad + Gem + Bu组合可将两种细胞系的增殖抑制至约55% - 60%。在此组合中添加SAHA可进一步将增殖降低至约30%。暴露于Clad + Gem + Bu + SAHA组合可激活DNA损伤反应和ATM - CHK2途径；修饰组蛋白；降低线粒体膜电位，导致凋亡诱导因子泄漏；并激活凋亡。用泛半胱天冬酶抑制剂Z - VAD - FMK预处理细胞可阻断组蛋白2AX的磷酸化以及PARP - 1和半胱天冬酶的裂解。Clad + Gem + Bu + SAHA组合在淋巴瘤细胞系中具有协同细胞毒性。我们的结果可能为在接受造血干细胞移植的淋巴瘤患者的临床试验中使用该组合作为移植前预处理方案提供依据，以替代更昂贵的核苷类似物氯法拉滨。
