Zhao Rui, Cui Yu, Li Dongbei, Guo Xiaoli, Cheng Cheng, He Rongheng, Hu Chenxi, Wei Xudong
Department of Hematopathy, Henan Institute of Hematology, The Affiliated Cancer Hospital of Zhengzhou University & Henan Cancer Hospital, Zhengzhou 450008, China.
Central Laboratory, The Affiliated Cancer Hospital of Zhengzhou University & Henan Cancer Hospital, Zhengzhou 450008, China.
Leuk Res. 2025 Feb;149:107637. doi: 10.1016/j.leukres.2024.107637. Epub 2024 Dec 20.
APG-115 is a novel small-molecule selective inhibitor that destabilizes the p53-MDM2 complex and activates p53-mediated apoptosis in tumor cells. Anlotinib inhibits tumor angiogenesis and promotes apoptosis. In this study, we investigated the apoptotic effect and potential mechanism of APG-115 and anlotinib combination on AML cell lines with different p53 backgrounds.
The IC50 values of APG-115 and anlotinib were detected by CCK-8 assay. The apoptosis rate of AML cells was evaluated by Annexin-V and PI double staining. Transcriptome sequencing was performed on the MOLM16 cell line treated with APG-115 and anlotinib, and differential analysis and enrichment analysis were performed. Real-time quantitative PCR and Western blot were used to detect the changes in cell cycle and pathway-related genes and proteins in AML cell lines after drug treatment. In vivo experiments, the anti-leukemia effects of APG-115 and anlotinib on AML xenograft mouse models were evaluated.
APG-115 and anlotinib could independently promote AML cell apoptosis, and the combination of the two drugs could produce a synergistic effect. Transcriptome sequencing showed that compared with the APG-115 monotherapy group, the differentially expressed genes were mainly enriched in the MDM2-p53 and PI3K/AKT pathways. In vivo experiments showed that compared with AML xenograft mice treated with either drug alone, AML progression was slowed in AML xenograft mice treated with APG-115 and anlotinib.
In vivo and in vitro experimental have shown that APG-115 combined with anlotinib can promote AML cells apoptosis and inhibit the progression of disease is independent of the p53 status.
APG-115是一种新型小分子选择性抑制剂，可使p53-MDM2复合物不稳定，并激活肿瘤细胞中p53介导的凋亡。安罗替尼可抑制肿瘤血管生成并促进凋亡。在本研究中，我们探究了APG-115与安罗替尼联合应用对不同p53背景的急性髓系白血病（AML）细胞系的凋亡作用及潜在机制。
采用CCK-8法检测APG-115和安罗替尼的半数抑制浓度（IC50）值。采用Annexin-V和碘化丙啶（PI）双染法评估AML细胞的凋亡率。对经APG-115和安罗替尼处理的MOLM16细胞系进行转录组测序，并进行差异分析和富集分析。采用实时定量聚合酶链反应（PCR）和蛋白质免疫印迹法检测药物处理后AML细胞系中细胞周期及通路相关基因和蛋白的变化。在体内实验中，评估APG-115和安罗替尼对AML异种移植小鼠模型的抗白血病作用。
APG-115和安罗替尼均可独立促进AML细胞凋亡，二者联合应用可产生协同效应。转录组测序显示，与APG-115单药治疗组相比，差异表达基因主要富集于MDM2-p53和磷脂酰肌醇-3-激酶（PI3K）/蛋白激酶B（AKT）通路。体内实验显示，与单独使用任一药物治疗的AML异种移植小鼠相比，联合使用APG-115和安罗替尼治疗的AML异种移植小鼠的疾病进展减缓。
体内和体外实验均表明，APG-115与安罗替尼联合应用可促进AML细胞凋亡并抑制疾病进展，且这一作用与p53状态无关。
