Immunology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Department of Cancer and Inflammation Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.
Curr Gene Ther. 2021;21(2):149-159. doi: 10.2174/1566523221999210120210017.
Acute myeloid leukemia (AML) is the most prevalent type of cancer in the adult hematopoietic system. Conventional therapies are associated with unfavorable side effects in individuals diagnosed with AML. These after-effects with partial remission reflect the urgent need for novel therapeutic approaches for inducing apoptosis, specifically in malignant cells, without affecting other cells. As a transcription factor (TF), ZEB2 (Zinc Finger E-Box Binding Homeobox 2) regulates the expression of specific genes in normal conditions. However, increased expression of ZEB2 is reported in various cancers, especially in AML, which is related to a higher degree of apoptosis inhibition of malignant cells. In this work, the role of ZEB2 in apoptosis inhibition is surveyed through ZEB2 specific knocking-down in human myeloid leukemia HL-60 cells.
Transfection of HL-60 cells was conducted using ZEB2-siRNA at concentrations of 20, 40, 60, and 80 pmol within 24, 48, and 72 h. After determining the optimum dose and time, flow cytometry was used to measure the apoptosis rate. The MTT assay was also utilized to evaluate the cytotoxic impact of transfection on the cells. The expression of candidate genes was measured before and after transfection using qRT-PCR.
According to obtained results, suppression of ZEB2 expression through siRNA was associated with the induction of apoptosis, increased pro-apoptotic, and decreased anti-apoptotic gene expression. Transfection of ZEB2-siRNA was also associated with reduced cell proliferation and viability.
Our study results suggest that ZEB2 suppression in myeloid leukemia cells through apoptosis induction could be a proper therapeutic method.
急性髓细胞白血病（AML）是成人造血系统中最常见的癌症类型。传统疗法与 AML 患者的不良副作用相关。这些部分缓解后的后遗症反映出迫切需要新的治疗方法来诱导细胞凋亡，特别是在恶性细胞中，而不影响其他细胞。作为转录因子（TF），ZEB2（锌指 E-盒结合同源盒 2）在正常条件下调节特定基因的表达。然而，在各种癌症中，包括 AML，都报告了 ZEB2 的表达增加，这与恶性细胞凋亡抑制程度更高有关。在这项工作中，通过在人髓样白血病 HL-60 细胞中特异性敲低 ZEB2，研究了 ZEB2 在凋亡抑制中的作用。
在 24、48 和 72 小时内，将浓度为 20、40、60 和 80 pmol 的 ZEB2-siRNA 转染 HL-60 细胞。确定最佳剂量和时间后，使用流式细胞术测量细胞凋亡率。还使用 MTT 测定法评估转染对细胞的细胞毒性影响。使用 qRT-PCR 测量转染前后候选基因的表达。
根据获得的结果，通过 siRNA 抑制 ZEB2 表达与诱导细胞凋亡、增加促凋亡和减少抗凋亡基因表达有关。ZEB2-siRNA 的转染也与细胞增殖和活力降低有关。
我们的研究结果表明，通过诱导细胞凋亡抑制髓样白血病细胞中的 ZEB2 可能是一种适当的治疗方法。
