下调癌相关成纤维细胞外泌体来源的 miR-29b-1-5p 通过免疫球蛋白结构域包含蛋白 1/紧密连接蛋白 1 轴抑制血管生成拟态和胃癌细胞凋亡，同时加速其迁移和侵袭。
Downregulation of cancer-associated fibroblast exosome-derived miR-29b-1-5p restrains vasculogenic mimicry and apoptosis while accelerating migration and invasion of gastric cancer cells via immunoglobulin domain-containing 1/zonula occluden-1 axis.
机构信息
Endoscopy Center, Minhang Hospital, Fudan University, Shanghai, China.
出版信息
Cell Cycle. 2023 Sep;22(17):1807-1826. doi: 10.1080/15384101.2023.2231740. Epub 2023 Aug 16.
Cancer-associated fibroblast (CAF) exosomal miRNAs have gradually a hot spot in cancer therapy. This study mainly explores the effect of CAF-derived exosomal miR-29b-1-5p on gastric cancer (GC) cells. CAFs and exosomes were identified by Western blot and transmission electron microscopy. CAF-derived exosomes-GC cells co-culture systems were constructed. Effects of CAF-derived exosomal miR-29b-1-5p on GC cells were determined by cell counting kit-8, flow cytometry, wound healing, Transwell assays and Western blot. The relationship between miR-29b-1-5p and immunoglobulin domain-containing 1 (VSIG1) was assessed by TargetScan, dual-luciferase reporter and RNA immunoprecipitation (RIP) experiments. The interaction between VSIG1 and zonula occluden-1 (ZO-1) was detected by co-immunoprecipitation. Expressions of miR-29b-1-5p, VSIG1 and ZO-1 were determined by quantitative real-time PCR. Vascular mimicry (VM) was detected using immunohistochemistry and tube formation assays. Rescue experiments and xenograft tumor assays were used to further determine the effect of CAF-derived exosomal miR-29b-1-5p/VSIG1 on GC. VM structure, upregulation of miR-29b-1-5p, and downregulation of VSIG1 and ZO-1 were shown in GC tissues. MiR-29b-1-5p targeted VSIG1, which interacted with ZO-1. CAF-derived exosomal miR-29b-1-5p inhibitor suppressed the viability, migration, invasion and VM formation, but promoted the apoptosis of GC cells. MiR-29b-1-5p inhibitor increased levels of VSIG1, ZO-1 and E-cadherin, whilst decreasing levels of VE-cadherin, N-cadherin and Vimentin in vitro and in vivo, which however was partially reversed by shVSIG1. Downregulation of CAF-derived exosomal miR-29b-1-5p impeded GC tumorigenesis and VM structure in vivo by upregulating VSIG1/ZO-1 expression. Downregulation of CAF-derived exosomal miR-29b-1-5p inhibits GC progression via VSIG1/ZO-1 axis.
癌症相关成纤维细胞（CAF）外泌体 miRNA 逐渐成为癌症治疗的热点。本研究主要探讨 CAF 来源的外泌体 miR-29b-1-5p 对胃癌（GC）细胞的影响。通过 Western blot 和透射电镜鉴定 CAFs 和外泌体。构建 CAF 衍生的外泌体-GC 细胞共培养系统。通过细胞计数试剂盒-8、流式细胞术、划痕愈合、Transwell 测定和 Western blot 测定确定 CAF 衍生的外泌体 miR-29b-1-5p 对 GC 细胞的影响。通过靶标扫描、双荧光素酶报告和 RNA 免疫沉淀（RIP）实验评估 miR-29b-1-5p 与免疫球蛋白结构域包含 1（VSIG1）的关系。通过共免疫沉淀检测 VSIG1 与紧密连接蛋白-1（ZO-1）的相互作用。通过定量实时 PCR 测定 miR-29b-1-5p、VSIG1 和 ZO-1 的表达。通过免疫组织化学和管形成测定检测血管模拟（VM）。通过拯救实验和异种移植肿瘤实验进一步确定 CAF 衍生的外泌体 miR-29b-1-5p/VSIG1 对 GC 的影响。GC 组织中可见 VM 结构、miR-29b-1-5p 上调、VSIG1 和 ZO-1 下调。miR-29b-1-5p 靶向 VSIG1，与 ZO-1 相互作用。CAF 衍生的外泌体 miR-29b-1-5p 抑制剂抑制 GC 细胞的活力、迁移、侵袭和 VM 形成，但促进 GC 细胞凋亡。miR-29b-1-5p 抑制剂在体外和体内增加 VSIG1、ZO-1 和 E-钙粘蛋白的水平，同时降低 VE-钙粘蛋白、N-钙粘蛋白和波形蛋白的水平，但通过 shVSIG1 部分逆转。下调 CAF 衍生的外泌体 miR-29b-1-5p 通过上调 VSIG1/ZO-1 表达抑制体内 GC 肿瘤发生和 VM 结构。下调 CAF 衍生的外泌体 miR-29b-1-5p 通过 VSIG1/ZO-1 轴抑制 GC 进展。
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