Department of Health Medicine, Second Medical Center of Chinese PLA General Hospital, No.28, Fuxing Road, Beijing, 100853, China.
Key Laboratory of Animal Ecology and Conservation Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing, China.
Dig Dis Sci. 2021 Dec;66(12):4274-4289. doi: 10.1007/s10620-020-06816-z. Epub 2021 Jan 15.
Exosome-mediated transfer of circular RNAs (circRNAs) is related to gastric cancer (GC) development. CircRNA NIMA-related kinase 9 (circNEK9; hsa_circ_0032683) was reported to be up-regulated in GC.
The biological role of circNEK9 and its underlying mechanisms in GC progression were explored.
The levels of RNAs and proteins were determined by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot assay. Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, colony formation assay, and flow cytometry. Wound healing assay and transwell assays were conducted to analyze cell motility. Intermolecular interaction was verified by dual-luciferase reporter assay and RNA pull-down assay. Animal experiments were used to evaluate the role of circNEK9 in the growth of xenograft tumors in vivo.
CircNEK9 was up-regulated in GC tissues and cell lines. CircNEK9 interference suppressed the proliferation and motility of GC cells. CircNEK9 silencing enhanced microRNA-409-3p (miR-409-3p) level through direct interaction. CircNEK9 silencing-mediated influences on the proliferation and metastasis of GC cells were partly overturned by the interference of miR-409-3p. MiR-409-3p directly interacted with microtubule-associated protein 7 (MAP7) messenger RNA (mRNA). MiR-409-3p-induced effects in GC cells were largely counteracted by the overexpression of MAP7. CircNEK9 silencing blocked GC tumor growth in vivo. Exosome-mediated transfer of circNEK9 promoted the motility of recipient GC cells.
CircNEK9 accelerated the proliferation, migration, and invasion of GC cells through targeting miR-409-3p/MAP7 axis. Plasma exosomal circNEK9 promoted the migration and invasion of recipient GC cells.
外泌体介导的环状 RNA（circRNA）转移与胃癌（GC）的发展有关。环状 RNA NIMA 相关激酶 9（circNEK9；hsa_circ_0032683）在 GC 中被报道上调。
探讨 circNEK9 在 GC 进展中的生物学作用及其潜在机制。
通过实时定量聚合酶链反应（qRT-PCR）和 Western blot 检测 RNA 和蛋白质水平。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）检测、集落形成实验和流式细胞术评估细胞增殖。通过划痕愈合实验和 Transwell 实验分析细胞迁移能力。通过双荧光素酶报告基因实验和 RNA 下拉实验验证分子间相互作用。通过动物实验评估 circNEK9 在体内异种移植肿瘤生长中的作用。
circNEK9 在 GC 组织和细胞系中上调。circNEK9 干扰抑制 GC 细胞的增殖和迁移。circNEK9 沉默通过直接相互作用增强了 microRNA-409-3p（miR-409-3p）水平。circNEK9 沉默介导的 GC 细胞增殖和转移的影响部分被 miR-409-3p 的干扰所逆转。miR-409-3p 与微管相关蛋白 7（MAP7）信使 RNA（mRNA）直接相互作用。miR-409-3p 在 GC 细胞中的作用在很大程度上被 MAP7 的过表达所抵消。circNEK9 沉默抑制体内 GC 肿瘤生长。外泌体介导的 circNEK9 转移促进了受体 GC 细胞的迁移。
circNEK9 通过靶向 miR-409-3p/MAP7 轴加速 GC 细胞的增殖、迁移和侵袭。血浆外泌体 circNEK9 促进了受体 GC 细胞的迁移和侵袭。
