Zhuang Yan, Zhao Jing, Xu Xi, Bi Laixi
Department of Hematology, the First Affiliated Hospital of Wenzhou Medical University, Wenzhou 325000, Zhejiang Province, China.
Department of Ophthalmology, Wenzhou City Hospital of Traditional Chinese Medicine and Western Medicine Combined, Wenzhou 325000, Zhejiang Province, China.
Arch Iran Med. 2018 Feb 1;21(2):73-78.
Glucose transporter 3 (GLUT3) plays an important role in tumor progression and drug resistance in numerous malignancies, including acute myeloid leukemia (AML). However, the effect of GLUT3 silencing on treatment of AML remains poorly understood. The purpose of this study was to investigate role of GLUT3 in proliferation and chemosensitivity of AML and its underlying mechanisms.
The siRNA transfection was conducted using LipofectamineTM 2000. Quantitative real-time RT-PCR (qRT-PCR) and Western blot analyses were employed to measure the expression levels of mRNA and protein for GLUT3, respectively. The cytotoxic effects of siRNA and vincristine were determined using the MTT assay. Flow cytometry was performed to analyze apoptosis.
GLUT3 siRNA transfection significantly reduced expression levels of GLUT3 mRNA and protein, leading to a strong growth inhibition and enhanced apoptosis (P = 0.017) in AML cells. Moreover, treatment with GLUT3 siRNA, synergistically enhanced the cytotoxic and apoptotic effects of vincristine (P = 0.025). We further investigated the possible mechanism involved in regulation of GLUT3 in AML cell proliferation and apoptosis. We found that GLUT3 negatively regulates EGFR activity, as well as the expression of its downstream proteins.
Our results demonstrated that GLUT3 plays a fundamental role in the survival and resistance of AML cells to vincristine. Therefore, GLUT3 can be considered as an attractive target for gene therapy of AML patients and siRNA-mediated silencing of this gene may be a novel strategy in AML treatment.
葡萄糖转运蛋白3（GLUT3）在包括急性髓系白血病（AML）在内的多种恶性肿瘤的肿瘤进展和耐药性中发挥重要作用。然而，GLUT3沉默对AML治疗的影响仍知之甚少。本研究的目的是探讨GLUT3在AML增殖和化疗敏感性中的作用及其潜在机制。
使用LipofectamineTM 2000进行siRNA转染。采用定量实时RT-PCR（qRT-PCR）和蛋白质印迹分析分别检测GLUT3的mRNA和蛋白质表达水平。使用MTT法测定siRNA和长春新碱的细胞毒性作用。进行流式细胞术分析细胞凋亡。
GLUT3 siRNA转染显著降低了GLUT3 mRNA和蛋白质的表达水平，导致AML细胞强烈的生长抑制和凋亡增强（P = 0.017）。此外，GLUT3 siRNA处理协同增强了长春新碱的细胞毒性和凋亡作用（P = 0.025）。我们进一步研究了GLUT3在AML细胞增殖和凋亡调控中的可能机制。我们发现GLUT3负向调节EGFR活性及其下游蛋白的表达。
我们的结果表明GLUT3在AML细胞对长春新碱的存活和耐药中起重要作用。因此，GLUT3可被视为AML患者基因治疗的有吸引力的靶点，并且该基因的siRNA介导的沉默可能是AML治疗的一种新策略。
