Exosomes-derived miR-154-5p attenuates esophageal squamous cell carcinoma progression and angiogenesis by targeting kinesin family member 14

Exosomes participate in the progression and angiogenesis of esophageal squamous cell carcinoma (ESCC). This study aimed to explore the effect and mechanism of exosomes-derived miR-154-5p on the progression and angiogenesis of ESCC. The exosomes with the diameter of 40–270 nm were successfully isolated from ESCC cells by ultracentrifugation. They were then assessed by transmission electron microscope (TEM), nanoparticle tracking analysis (NTA), and Western blotting. Kinesin family member 14 (KIF14) was upregulated, while miR-154-5p was downregulated in ESCC as examined by Quantitative Real-time PCR (qRT-PCR). Exosomes-derived miR-154-5p from ESCC cells was found to attenuate the cellular migration, invasion, and angiogenesis of ESCC using Cell Counting Kit-8 (CCK-8), wound healing assay, transwell migration assay, and tumor formation assays. Moreover, KIF14 was proven to be a direct downstream target gene of miR-154-5p in ESCC cells using luciferase assay. In conclusion, our study identified that exosomes-derived miR-154-5p attenuates ESCC progression and angiogenesis by targeting KIF14 <i>in vitro</i>, which might provide a novel approach for the diagnosis and treatment of ESCC.

外泌体（Exosomes）参与食管鳞状细胞癌（esophageal squamous cell carcinoma, ESCC）的进展与血管生成过程。本研究旨在探讨外泌体来源的微小RNA-154-5p（miR-154-5p）对ESCC进展及血管生成的作用与机制。本研究通过超速离心法从ESCC细胞中成功分离得到直径为40~270 nm的外泌体，并通过透射电子显微镜（transmission electron microscope, TEM）、纳米颗粒追踪分析（nanoparticle tracking analysis, NTA）以及蛋白质印迹（Western blotting）对其进行鉴定。经实时荧光定量聚合酶链反应（Quantitative Real-time PCR, qRT-PCR）检测发现，ESCC细胞中驱动蛋白家族成员14（Kinesin family member 14, KIF14）表达上调，而miR-154-5p表达下调。通过细胞计数试剂盒-8（Cell Counting Kit-8, CCK-8）、划痕愈合实验、Transwell迁移实验及成瘤实验证实，ESCC细胞来源的外泌体miR-154-5p可减弱ESCC细胞的迁移、侵袭能力及血管生成能力。此外，本研究通过荧光素酶报告基因实验（luciferase assay）证实，KIF14是ESCC细胞中miR-154-5p的直接下游靶基因。综上，本研究明确了外泌体来源的miR-154-5p可通过靶向KIF14在体外（in vitro）抑制ESCC的进展与血管生成，该发现可为ESCC的诊断与治疗提供全新策略。