Departments of Gastrointestinal Surgery, The First Affiliated Hospital of Guangxi Medical University, 530021, Nanning, P.R. China.
Guangxi Key Laboratory of Enhanced Recovery after Surgery for Gastrointestinal Cancer, 530021, Nanning, P.R. China.
Cell Death Dis. 2022 Mar 8;13(3):217. doi: 10.1038/s41419-021-04446-5.
Gastric cancer (GC) is a heterogeneous disease with poor prognosis. Tumor-derived extracellular vesicles (EVs) assume a role in intercellular communication by carrying various molecules, including proteins, RNA, and DNAs, which has been identified to exhibit oncogenic effect in GC. Therefore, this research aimed to figure out whether tumor-derived EVs transmit c-Myc to orchestrate the growth and metastasis of GC. KCNQ1OT1, microRNA (miR)-556-3p and CLIC1 expression of GC tissues was detected through RT-qPCR. EVs were isolated from GC cells, followed by RT-qPCR and Western blot analysis of c-Myc expression in EVs and GC cells. Next, GC cells were incubated with EVs or transfected with a series of mimic, inhibitor, or siRNAs to assess their effects on cell viability, migrative, invasive, and apoptotic potential. Relationship among c-Myc, KCNQ1OT1, miR-556-3p, and CLIC1 was evaluated by dual-luciferase reporter assay. PI3K/AKT pathway-related proteins were assessed through Western blot analysis. KCNQ1OT1 and CLIC1 were highly expressed but miR-556-3p in GC tissues. c-Myc was high-expressed in tumor-derived EVs and GC cells. Mechanistically, c-Myc could induce KCNQ1OT1 expression, and KCNQ1OT1 bound to miR-556-3p that negatively targeted CLIC1 to inactivate PI3K/AKT pathway. Tumor-derived EVs, EVs-c-Myc, KCNQ1OT1 or CLIC1 overexpression, or miR-556-3p inhibition promoted GC cell proliferative, invasive, and migrative capacities but repressed their apoptosis through activating PI3K/AKT pathway. Collectively, tumor-derived EVs carrying c-Myc activated KCNQ1OT1 to downregulate miR-556-3p, thus elevating CLIC1 expression to activate the PI3K/AKT pathway, which facilitated the growth and metastasis of GC.
胃癌(GC)是一种预后不良的异质性疾病。肿瘤衍生的细胞外囊泡(EVs)通过携带各种分子(包括蛋白质、RNA 和 DNA)在细胞间通讯中发挥作用,这些分子已被确定在 GC 中具有致癌作用。因此,本研究旨在探讨肿瘤衍生的 EV 是否通过传递 c-Myc 来协调 GC 的生长和转移。通过 RT-qPCR 检测 GC 组织中的 KCNQ1OT1、microRNA(miR)-556-3p 和 CLIC1 的表达。从 GC 细胞中分离 EVs,然后通过 RT-qPCR 和 Western blot 分析 EVs 和 GC 细胞中 c-Myc 的表达。接下来,将 GC 细胞与 EVs 孵育或用一系列模拟物、抑制剂或 siRNA 转染,以评估它们对细胞活力、迁移、侵袭和凋亡潜力的影响。通过双荧光素酶报告基因检测评估 c-Myc、KCNQ1OT1、miR-556-3p 和 CLIC1 之间的关系。通过 Western blot 分析评估 PI3K/AKT 通路相关蛋白。在 GC 组织中,KCNQ1OT1 和 CLIC1 高表达,但 miR-556-3p 低表达。c-Myc 在肿瘤衍生的 EVs 和 GC 细胞中高表达。机制上,c-Myc 可以诱导 KCNQ1OT1 的表达,KCNQ1OT1 与负靶向 CLIC1 的 miR-556-3p 结合,从而使 PI3K/AKT 通路失活。肿瘤衍生的 EVs、EVs-c-Myc、KCNQ1OT1 或 CLIC1 的过表达或 miR-556-3p 的抑制通过激活 PI3K/AKT 通路促进 GC 细胞的增殖、侵袭和迁移能力,但抑制其凋亡。综上所述,携带 c-Myc 的肿瘤衍生 EV 通过激活 KCNQ1OT1 下调 miR-556-3p,从而升高 CLIC1 表达激活 PI3K/AKT 通路,促进 GC 的生长和转移。