Zahed Panah Mahdi, Nikbakht Mohsen, Sajjadi Seyed Mehdi, Rostami Shahrbano, Norooznezhad Amir Hossein, Kamranzadeh Fumani Hosein, Ghavamzadeh Ardeshir, Mohammadi Saeed
Department of Medical Laboratory Sciences, Faculty of Allied Medicine, Qazvin University of Medical Sciences, Qazvin, Iran.
Hematology-Oncology and Stem Cell Transplantation Research Center, Tehran University of Medical Sciences, Tehran, Iran.
Int J Hematol Oncol Stem Cell Res. 2017 Apr 1;11(2):148-157.
The conventional chemotherapeutic regimens which applied for treatment of acute myeloid leukemia (AML) mostly target tumor bulk but not leukemic stem cells (LSCs). Aberrant expression or activation of mediators such as osteopontin (OPN) or PI3K/PTEN/Akt/mTOR pathway plays a key role in making prone to develop leukemia. Preventing or treating cancer by curcumin (CUR) has been suggested recently. CUR induces apoptosis and growth inhibition through various mechanisms in leukemic cells. In present study, we tried to measure the toxic response in vitro to CUR for evaluation ofchangesin cell viability, survival and molecular-mediated resistance in primary AML cells. Isolated primary CD34+/CD38- bone marrow derived AML cells were treated with CUR, Daunorubicin (DNR) and/or their combination by MTT assay, Annexin V/PI staining, and colony-formation. The mRNA expression of OPN/AKT/mTOR/PTEN/β-catenin genes was measured by Real-Time PCR. The siRNA against OPN was applied for CUR- treated cells. Growth inhibition effect of DNR increased in combination with CUR on primary CD34+/CD38- AML cells. Suppression of OPN with siRNA increased the cytotoxic effects of CUR. Likewise, OPN gene expression increased in response to CUR treatment in AML cells. AKT, mTOR, β-catenin or PTEN gene expression increased by CUR, but OPN siRNA decreased the level of mRNA expression of mentioned molecular pathway. The chemo-resistance of AML cells against therapy might be relevant to increasing of OPN mRNA expression and activity of other mediators including AKT, mTOR, PTEN, and β-catenin. In this context, targeting of OPN might be more impact on CD34+ AML cells.
用于治疗急性髓系白血病（AML）的传统化疗方案大多针对肿瘤细胞团，而非白血病干细胞（LSCs）。骨桥蛋白（OPN）或PI3K/PTEN/Akt/mTOR等信号分子的异常表达或激活在白血病的发生发展中起关键作用。近期有研究表明姜黄素（CUR）可预防或治疗癌症。CUR通过多种机制诱导白血病细胞凋亡并抑制其生长。在本研究中，我们试图检测原发性AML细胞对CUR的体外毒性反应，以评估细胞活力、存活率及分子介导的耐药性变化。采用MTT法、Annexin V/PI染色法和集落形成法，用CUR、柔红霉素（DNR）及其联合用药处理分离的原发性CD34+/CD38-骨髓来源的AML细胞。通过实时定量PCR检测OPN/AKT/mTOR/PTEN/β-连环蛋白基因的mRNA表达。将针对OPN的小干扰RNA（siRNA）应用于CUR处理的细胞。DNR与CUR联合使用时，对原发性CD34+/CD38- AML细胞的生长抑制作用增强。用siRNA抑制OPN可增强CUR的细胞毒性作用。同样，AML细胞经CUR处理后OPN基因表达增加。CUR可使AKT、mTOR、β-连环蛋白或PTEN基因表达增加，但OPN siRNA可降低上述分子通路的mRNA表达水平。AML细胞对化疗的耐药性可能与OPN mRNA表达增加以及AKT、mTOR、PTEN和β-连环蛋白等其他信号分子的活性增强有关。在这种情况下，靶向OPN可能对CD34+ AML细胞有更大影响。
