Shanghai Institute of Hematology, State Key Laboratory of Medical Genomics, National Research Center for Translational Medicine at Shanghai, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of General Practice, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
PLoS One. 2024 Apr 9;19(4):e0299019. doi: 10.1371/journal.pone.0299019. eCollection 2024.
Multiple myeloma (MM) is the second most prevalent hematologic malignancy which remains uncurable. Numerous drugs have been discovered to inhibit MM cells. Indisulam, an aryl sulfonamide, has a potent anti-myeloma activity in vitro and in vivo. This study aims to explore the new mechanism of indisulam and investigate its potential use in combination with melphalan. We examined DNA damage in MM cells through various methods such as western blotting (WB), immunofluorescence, and comet assay. We also identified the role of topoisomerase IIα (TOP2A) using bioinformatic analyses. The impact of indisulam on the RNA and protein levels of TOP2A was investigated through qPCR and WB. Cell proliferation and apoptosis were assessed using CCK-8 assays, Annexin V/PI assays and WB. We predicted the synergistic effect of the combination treatment based on calculations performed on a website, and further explored the effect of indisulam in combination with melphalan on MM cell lines and xenografts. RNA sequencing data and basic experiments indicated that indisulam caused DNA damage and inhibited TOP2A expression by decreasing transcription and promoting degradation via the proteasome pathway. Functional experiments revealed that silencing TOP2A inhibited cell proliferation and induced apoptosis and DNA damage. Finally, Indisulam/melphalan combination treatment demonstrated a strong synergistic anti-tumor effect compared to single-agent treatments in vitro and in vivo. These findings suggest that combination therapies incorporating indisulam and melphalan have the potential to enhance treatment outcomes for MM.
多发性骨髓瘤(MM)是第二大常见的血液系统恶性肿瘤,目前仍然无法治愈。已经发现许多药物可以抑制 MM 细胞。芳基磺酰胺类药物 indisulam 在体外和体内均具有很强的抗骨髓瘤活性。本研究旨在探索 indisulam 的新作用机制,并研究其与美法仑联合应用的潜力。我们通过 Western blot(WB)、免疫荧光和彗星试验等多种方法检测 MM 细胞中的 DNA 损伤。我们还通过生物信息学分析鉴定了拓扑异构酶 IIα(TOP2A)的作用。通过 qPCR 和 WB 研究了 indisulam 对 TOP2A 的 RNA 和蛋白水平的影响。通过 CCK-8 测定、Annexin V/PI 测定和 WB 评估细胞增殖和凋亡。我们根据网站上的计算预测联合治疗的协同作用,并进一步探索 indisulam 与美法仑联合治疗对 MM 细胞系和异种移植物的影响。RNA 测序数据和基础实验表明,indisulam 通过降低转录和促进通过蛋白酶体途径降解来引起 DNA 损伤并抑制 TOP2A 表达。功能实验表明,沉默 TOP2A 抑制细胞增殖并诱导细胞凋亡和 DNA 损伤。最后,与单药治疗相比,indisulam/美法仑联合治疗在体外和体内均显示出强烈的协同抗肿瘤作用。这些发现表明,包含 indisulam 和美法仑的联合治疗可能增强 MM 的治疗效果。