Song Ying-Hui, Zhong Mei-Zuo, Gan Ping-Ping, Yi Ping-Yong, Tang You-Hong, Liu Yi-Ping, Jiang Jin-Qiong, Li Li
Department of Oncology, Xiangya Hospital, Central South University, 88 Xiangya Road, Changsha, 410008, Hunan, People's Republic of China.
Tumour Biol. 2014 Dec;35(12):11809-17. doi: 10.1007/s13277-014-2335-9. Epub 2014 Oct 25.
Although there have been substantial advances in our knowledge of the resistance of diffuse large B cell lymphoma (DLBCL) to chemotherapy, there are few efficient treatment strategies for recurrent/refractory DLBCL. The aim of this study was to investigate the role of aldehyde dehydrogenase (ALDH) 1A1 in the resistance of diffuse large B cell lymphoma to the chemotherapeutic mixture consisting of cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP). The involvement of ALDH1A1 in DLBCL was elucidated by knockdown and pharmacologic inhibition; Cell Counting Kit-8 (CCK-8) and clone formation assays were used to determine its role in CHOP sensitivity and clone formation ability. Caspase colorimetric assay was used to measure the extent of apoptosis. Western blot analysis was used to measure signal transducer and activator of transcription 3 (STAT3)/nuclear factor kappa B (NF-κB) signaling proteins, and quantitative real-time PCR (RT-PCR) was used to measure the differential expression of ALDH1A1 of DLBCL patients and healthy donors. ALDH1A1 showed a 5.64-fold higher expression in malignant B cells than in normal B cells. Diethylaminobenzaldehyde (DEAB) decreased the half maximal inhibitory concentration (IC50) of the CHOP regimen in Farage cells from 344.78 ± 65.75 to 183.88 ± 49.75 ng/ml (P = 0.004). Both knockdown and inhibition of ALDH1A1 reduced clonogenicity, increased caspase-3/caspase-9 activity, and attenuated the phosphorylation status of STAT3/NF-κB. The prognosis of patients with a high level of ALDH1A1 expression was poor compared with that of patients with low levels of expression (P = 0.044). ALDH1A1 is a new mediator for resistance of DLBCL to CHOP; it is a predictor of clinical prognosis and may serve as a potential target to improve chemotherapy responsiveness of human DLBCL.
尽管我们对弥漫性大B细胞淋巴瘤（DLBCL）对化疗的耐药性已有了实质性进展，但对于复发/难治性DLBCL，仍缺乏有效的治疗策略。本研究旨在探讨醛脱氢酶（ALDH）1A1在弥漫性大B细胞淋巴瘤对由环磷酰胺、阿霉素、长春新碱和泼尼松组成的化疗方案（CHOP）耐药中的作用。通过敲低和药物抑制来阐明ALDH1A1在DLBCL中的作用；使用细胞计数试剂盒-8（CCK-8）和克隆形成试验来确定其在CHOP敏感性和克隆形成能力中的作用。使用半胱天冬酶比色法测量凋亡程度。蛋白质免疫印迹分析用于测量信号转导和转录激活因子3（STAT3）/核因子κB（NF-κB）信号蛋白，定量实时聚合酶链反应（RT-PCR）用于测量DLBCL患者和健康供体中ALDH1A1的差异表达。ALDH1A1在恶性B细胞中的表达比正常B细胞高5.64倍。二乙氨基苯甲醛（DEAB）使Farage细胞中CHOP方案的半数最大抑制浓度（IC50）从344.78±65.75降至183.88±49.75 ng/ml（P = 0.004）。敲低和抑制ALDH1A1均降低了克隆形成能力，增加了半胱天冬酶-3/半胱天冬酶-9活性，并减弱了STAT3/NF-κB的磷酸化状态。与低表达水平的患者相比，ALDH1A1表达水平高的患者预后较差（P = 0.044）。ALDH1A1是DLBCL对CHOP耐药的新介质；它是临床预后的预测指标，可能作为改善人类DLBCL化疗反应性的潜在靶点。
