Wang Xiao-Bo, Yuan Li-Hua, Yan Le-Ping, Ye Yong-Bin, Lu Bo, Xu Xiaojun
Department of Hematology, The Seventh Affiliated Hospital, Sun Yat-Sen University, Shenzhen, China.
Department of Pediatric Surgery, University of Hong Kong-Shenzhen Hospital, Shenzhen, China.
Front Oncol. 2022 May 30;12:920999. doi: 10.3389/fonc.2022.920999. eCollection 2022.
In clinical practice, arsenic trioxide can be used to treat a subset of R/R CML patients, but resistance tends to reappear quickly. We designed an experiment to study arsenic trioxide resistance in K-562 cells. Previously, we identified the gene as potentially responsible for arsenic trioxide resistance in K-562 cells gene chip screening followed by high-content screening. We aimed to investigate the role and mechanism of the gene in K-562 cells, an arsenic trioxide-resistant chronic myeloid leukemia cell line. lentiviral vector-mediated siRNA transfection was performed on K-562 cells. The roles of in cell proliferation, apoptosis and cell cycle pathways, and colony formation were analyzed by CCK-8 assay, fluorescence-activated cell sorting, and soft agar culture, respectively. Gene chip screening was used to define the possible downstream pathways of . Western blot was performed to further validate the possible genes mediated by for arsenic trioxide resistance in patients with chronic myeloid leukemia. downregulation significantly inhibited growth, promoted apoptosis, decreased colony formation, reduced the duration of the G1 phase, and increased the duration of the S phase of K-562 cells. Western blot results confirmed that may modulate the apoptosis and proliferation of arsenic trioxide-resistant chronic myeloid leukemia cells through the mediation of MAP3K7, CDK4, and PINK1. is a potential therapeutic target for patients with arsenic trioxide-resistant chronic myeloid leukemia.
在临床实践中，三氧化二砷可用于治疗一部分复发/难治性慢性髓性白血病（R/R CML）患者，但耐药性往往很快再次出现。我们设计了一项实验来研究K-562细胞中的三氧化二砷耐药性。此前，我们通过基因芯片筛选和高内涵筛选，确定该基因可能是K-562细胞中三氧化二砷耐药性的原因。我们旨在研究该基因在K-562细胞（一种耐三氧化二砷的慢性髓性白血病细胞系）中的作用和机制。对K-562细胞进行慢病毒载体介导的siRNA转染。分别通过CCK-8检测、荧光激活细胞分选和软琼脂培养分析该基因在细胞增殖、凋亡和细胞周期途径以及集落形成中的作用。利用基因芯片筛选来确定该基因可能的下游途径。进行蛋白质免疫印迹法以进一步验证慢性髓性白血病患者中该基因介导的三氧化二砷耐药性可能相关的基因。该基因的下调显著抑制了K-562细胞的生长，促进了凋亡，减少了集落形成，缩短了G1期的持续时间，并增加了S期的持续时间。蛋白质免疫印迹结果证实，该基因可能通过介导MAP3K7、CDK4和PINK1来调节耐三氧化二砷的慢性髓性白血病细胞的凋亡和增殖。该基因是耐三氧化二砷的慢性髓性白血病患者的一个潜在治疗靶点。
