Han Hu, Li Yan, Lin Zhiyi, Ma Xiaoping, Huang Wukui, Lu Cengceng, Ma Rongyan, Han Rui
Department of Oncology, The First Affiliated Hospital of Shihezi University, Shihezi 832008, China.
Medical Department, The First Affiliated Hospital of Shihezi University, Shihezi 832008, China.
Int Immunopharmacol. 2025 Jan 27;146:113804. doi: 10.1016/j.intimp.2024.113804. Epub 2024 Dec 16.
Exosomal microRNA (miRNA)s have been proven to affect recipient cell chemoresistance in several cancers. This research attempted to uncover the role of exosomal miRNA and the regulatory mechanism in cisplatin resistance of esophageal cancer (EC). Cisplatin-resistant EC cells (KYSE-150-CisR and TE-1-CisR) were established by the parental cells (KYSE-150 and TE-1) treated with a gradual increase of cisplatin concentration. Exosomes from both cisplatin-resistant EC cells and the parental one were obtained with ultracentrifugation (CisR-exo and CisS-exo), and identified by transmission electron microscopy and nanoparticle tracking analysis. The effect of CisR-exo on the cisplatin resistance of EC was assessed by in vitro (and in vivo functional experiments.Intracellular reactive oxygen species and iron were determined by the corresponding kits. The m6A dot blot assay and methylated RIP-qPCR was conducted to analyze the m6A modification level. Dual-luciferase reporter assay was performed to confirm the intermolecular interaction. Increased levels of miR-130a-3p were observed in both KYSE-150CisR and TE-1CisR cells, as well as their derived CisR-exos when compared with the parental cells and CisS-exos. Exosomal miR-130a-3p from cisplatin-resistant EC cells conferred cisplatin resistance to EC in vitro and in vivo, which might be mediated by the suppression of ferroptosis. Mechanically, KYSE-150CisR derived exosomal miR-130a-3p interacted with METTL14 to inhibit FSP1 (a ferroptosis suppressor) m6A modification of recipient cells. Overexpressing METTL14 restrained the cisplatin resistance disseminated by CisR-exos in KYSE-150 cells. Cisplatin-resistant EC cell-isolated exosomal miR-130a-3p suppressed the m6A modification of FSP1 to modulate ferroptosis, enhancing cisplatin resistance.
外泌体微小RNA（miRNA）已被证明在多种癌症中会影响受体细胞的化疗耐药性。本研究试图揭示外泌体miRNA在食管癌（EC）顺铂耐药中的作用及其调控机制。通过用逐渐增加浓度的顺铂处理亲代细胞（KYSE - 150和TE - 1）建立了顺铂耐药的EC细胞（KYSE - 150 - CisR和TE - 1 - CisR）。通过超速离心从顺铂耐药的EC细胞和亲代细胞中获得外泌体（CisR - exo和CisS - exo），并通过透射电子显微镜和纳米颗粒跟踪分析进行鉴定。通过体外和体内功能实验评估CisR - exo对EC顺铂耐药性的影响。使用相应试剂盒测定细胞内活性氧和铁的含量。进行m6A斑点印迹分析和甲基化RIP - qPCR以分析m6A修饰水平。进行双荧光素酶报告基因测定以确认分子间相互作用。与亲代细胞和CisS - exo相比，在KYSE - 150CisR和TE - 1CisR细胞及其衍生的CisR - exos中均观察到miR - 130a - 3p水平升高。来自顺铂耐药EC细胞的外泌体miR - 130a - 3p在体外和体内赋予EC顺铂耐药性，这可能是通过抑制铁死亡介导的。机制上，KYSE - 150CisR衍生的外泌体miR - 130a - 3p与METTL14相互作用，抑制受体细胞中FSP1（一种铁死亡抑制因子）的m6A修饰。过表达METTL14可抑制KYSE - 150细胞中CisR - exos传播的顺铂耐药性。顺铂耐药EC细胞分离出的外泌体miR - 130a - 3p抑制FSP1的m6A修饰以调节铁死亡，增强顺铂耐药性。
