Zhou Ling-Yun, Chen Fang-Yuan, Shen Li-Jing, Wan Hai-Xia, Zhong Ji-Hua
Department of Hematology, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200127, P.R. China.
Exp Ther Med. 2014 Jul;8(1):85-90. doi: 10.3892/etm.2014.1716. Epub 2014 May 15.
Acute leukemia is a malignant clonal hematopoietic stem cell disease. In the current study, the effects of arsenic trioxide (ATO) on the ecotropic viral integration site-1 (EVI-1) gene were investigated in the THP1 cell line. THP-1 cells were treated with different concentrations of ATO (0, 1, 3 and 5 μM) for 24, 48 or 72 h, then tested for cell viability by CCK-8 kit, cell morphology by cytospin smear, cell apoptosis by flow cytometry, EVI-1 mRNA expression by reverse transcription polymerase chain reaction (RT-PCR) and protein quantity by western blot. ATO treatment was shown to inhibit proliferation and induce apoptosis in THP1 cells in a dose- and time-dependent manner. ATO downregulated the mRNA and protein expression of EVI-1 in the THP1 cell line. In addition, ATO significantly decreased the expression of antiapoptotic proteins, B-cell lymphoma 2 (Bcl-2) and B cell lymphoma-extra large (Bcl-xL), but markedly increased the expression of proapoptotic proteins, including c-Jun N-terminal kinase (JNK), phosphorylated-JNK, Bax, full length caspase-3 and cleaved caspase-3. These results indicated that ATO inhibited the proliferation and induced apoptosis in THP1 cells partially via blocking the inhibitory effects of EVI-1 on the JNK signaling pathway with the involvement of apoptosis-associated proteins, including Bax, Bcl-2, Bcl-xL and caspase-3. These novel observations may be used to elucidate the mechanism by which ATO induces apoptosis in acute leukemia cells, and provide rationales to develop a personalized medicine strategy for ATO via targeting EVI-1 positive neoplasm.
急性白血病是一种恶性克隆性造血干细胞疾病。在本研究中，研究了三氧化二砷（ATO）对嗜亲性病毒整合位点1（EVI-1）基因在THP1细胞系中的影响。用不同浓度的ATO（0、1、3和5 μM）处理THP-1细胞24、48或72小时，然后通过CCK-8试剂盒检测细胞活力，通过细胞离心涂片检测细胞形态，通过流式细胞术检测细胞凋亡，通过逆转录聚合酶链反应（RT-PCR）检测EVI-1 mRNA表达，通过蛋白质印迹检测蛋白质含量。结果表明，ATO以剂量和时间依赖性方式抑制THP1细胞的增殖并诱导其凋亡。ATO下调THP1细胞系中EVI-1的mRNA和蛋白质表达。此外，ATO显著降低抗凋亡蛋白B细胞淋巴瘤2（Bcl-2）和B细胞淋巴瘤-特大（Bcl-xL）的表达，但明显增加促凋亡蛋白的表达，包括c-Jun氨基末端激酶（JNK）、磷酸化-JNK、Bax、全长半胱天冬酶-3和裂解的半胱天冬酶-3。这些结果表明，ATO通过阻断EVI-1对JNK信号通路的抑制作用，并涉及包括Bax、Bcl-2、Bcl-xL和半胱天冬酶-3在内的凋亡相关蛋白，部分抑制THP1细胞的增殖并诱导其凋亡。这些新发现可用于阐明ATO诱导急性白血病细胞凋亡的机制，并为通过靶向EVI-1阳性肿瘤开发ATO的个性化药物策略提供理论依据。
