Dehghan-Nayeri Nasrin, Rezaei-Tavirani Mostafa, Omrani Mir Davood, Gharehbaghian Ahmad, Goudarzi Pour Kourosh, Eshghi Peyman
Proteomics Research Center, School of Allied Medical Sciences, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
Department of Medical Genetics, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
J Cell Commun Signal. 2017 Jun;11(2):137-145. doi: 10.1007/s12079-016-0357-3. Epub 2016 Oct 24.
Response to dexamethasone (DEXA), as a hallmark drug in the treatment of childhood acute lymphoblastic leukemia (ALL), is one of the pivotal prognostic factors in the prediction of outcome in ALL. Identification of predictive markers of chemoresistance is beneficial to selecting of the best therapeutic protocol with the lowest effect adverse. Hence, we aimed to find drug targets using the 2DE/MS proteomics study of a DEXA-resistant cell line (REH) as a model for poor DEXA responding patients before and after drug treatment. Using the proteomic methods, three differentially expressed proteins were detected, including voltage dependent anion channel 1 (VDAC1), sorting Nexin 3 (SNX3), and prefoldin subunit 6 (PFDN6). We observed low expression of three proteins after DEXA treatment in REH cells. We subsequently verified low expression of resulted proteins at the mRNA level using the quantitative PCR method. These proteins are promising proteins because of their important roles in drug resistance and regulation of apoptosis (VDAC1), protein trafficking (SNX3), and protein folding (PFDN6). Additionally, mRNA expression level of these proteins was assessed in 17 bone marrow samples from children with newly diagnosed ALL and 7 non-cancerous samples as controls. The results indicated that independent of the molecular subtypes of leukemia, mRNA expression of VDAC1, SNX3, and PFDN6 decreased in ALL samples compared with non-cancerous samples particularly in VDAC1 (p < 0.001). Additionally, mRNA expression of three proteins was also declined in high-risk samples compared with standard risk cases. These results demonstrated diagnostic and prognostic value of these proteins in childhood ALL. Furthermore, investigation of protein-protein interaction using STRING database indicated that these proteins involved in the signaling pathway of NR3C1 as dexamethasone target. In conclusion, our proteomic study in DEXA resistant leukemic cells revealed VDAC1, SNX3, and PFDN6 are promising proteins that might serve as potential biomarkers of prognosis and chemotherapy in childhood ALL.
地塞米松（DEXA）作为治疗儿童急性淋巴细胞白血病（ALL）的标志性药物，其反应是ALL预后预测的关键预后因素之一。识别化疗耐药的预测标志物有助于选择疗效最佳且副作用最小的治疗方案。因此，我们旨在通过对DEXA耐药细胞系（REH）进行二维凝胶电泳/质谱（2DE/MS）蛋白质组学研究，以药物治疗前后DEXA反应不佳的患者为模型来寻找药物靶点。采用蛋白质组学方法，检测到三种差异表达蛋白，包括电压依赖性阴离子通道1（VDAC1）、分选连接蛋白3（SNX3）和预折叠蛋白亚基6（PFDN6）。我们观察到DEXA处理后REH细胞中这三种蛋白表达降低。随后，我们使用定量PCR方法在mRNA水平验证了这些蛋白表达降低。这些蛋白具有重要意义，因为它们在耐药性和细胞凋亡调节（VDAC1）、蛋白质转运（SNX3）以及蛋白质折叠（PFDN6）方面发挥重要作用。此外，在17例新诊断ALL儿童的骨髓样本和7例作为对照的非癌样本中评估了这些蛋白的mRNA表达水平。结果表明，与非癌样本相比，ALL样本中VDAC1、SNX3和PFDN6的mRNA表达降低，与白血病分子亚型无关，尤其是VDAC1（p < 0.001）。此外，与标准风险病例相比，高风险样本中这三种蛋白的mRNA表达也下降。这些结果证明了这些蛋白在儿童ALL中的诊断和预后价值。此外，使用STRING数据库对蛋白质 - 蛋白质相互作用的研究表明，这些蛋白参与了作为地塞米松靶点的NR3C1信号通路。总之，我们对DEXA耐药白血病细胞的蛋白质组学研究表明，VDAC1、SNX3和PFDN6是有前景的蛋白，可能作为儿童ALL预后和化疗的潜在生物标志物。
