Effect of Gilteritinib, Metformin and Combination treatments on gene expression in MOLM13-RES cells

Here, we report that Metformin shows a striking synergistic effect with Gilteritinib in suppressing cell proliferation and promoting apoptosis and cell cycle arrest in multiple FLT3-ITD AML cell lines, including FLT3 TKI-resistant MOLM13 cells. Mechanistically, the combinational treatment synergistically suppresses Polo-like kinase 1 (PLK1) expression and phosphorylation of FLT3/STAT5/ERK/mTOR in MOLM13-RES cells. Intriguingly, our retrospective clinical analysis has unveiled a significant correlation between Metformin intake and improved survival rates among FLT3-ITD AML patients. Collectively, the cotreatment of Metformin and Gilteritinib shows robustly enhanced therapeutic efficacy in treating FLT3-mutated AML by synergistically suppressing PLK1 expression and phosphorylation of FLT3/STAT5/ERK/mTOR. Overall design: MOLM13-RES cells were cultured under standard conditions and were treated with DMSO, Gilteritinib, Metformin and Combination, respectively. Collect the four group cells after 24 hours treatment and extract total RNA. Use 10ug for next library preparation and sequencing.

本研究证实，二甲双胍（Metformin）与吉列替尼（Gilteritinib）在多种FLT3内部串联重复型急性髓系白血病（FLT3-ITD AML）细胞系（包括FLT3酪氨酸激酶抑制剂（FLT3 TKI）耐药的MOLM13细胞）中，可发挥显著协同效应，抑制细胞增殖、促进细胞凋亡并诱导细胞周期阻滞。从机制层面而言，联合给药可在MOLM13-RES细胞中协同抑制polo样激酶1（PLK1）的表达，以及FLT3/STAT5/ERK/mTOR的磷酸化水平。值得注意的是，本研究的回顾性临床分析显示，二甲双胍的摄入与FLT3-ITD AML患者的生存率提升存在显著相关性。综上，二甲双胍与吉列替尼的联合治疗可通过协同抑制PLK1表达及FLT3/STAT5/ERK/mTOR磷酸化，显著增强FLT3突变型急性髓系白血病的治疗疗效。整体实验设计如下：将MOLM13-RES细胞置于标准培养条件下培养，分别以二甲基亚砜（DMSO）、吉列替尼、二甲双胍及联合给药方案进行处理。给药24小时后收集四组细胞并提取总RNA，取10μg用于后续文库制备及测序。