Colorectal cancer cell-derived extracellular vesicles transfer miR-221-3p to promote endothelial cell angiogenesis via targeting suppressor of cytokine signaling 3
Colorectal cancer cell-derived extracellular vesicles transfer miR-221-3p to promote endothelial cell angiogenesis via targeting suppressor of cytokine signaling 3
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DOI：
10.1016/j.lfs.2021.119937
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发表时间：
2021-09-16
期刊：
影响因子：
6.1
通讯作者：
Babashah，Sadegh
中科院分区：
文献类型：
--
作者：
Dokhanchi，Maryam;Pakravan，Katayoon;Babashah，Sadegh
背景分泌型微小RNA（microRNAs，miRNAs）是一种很有前途的结直肠癌（colorectal cancer，CRC）诊断标志物。在此，我们评估了四种循环血清来源的miRNA在CRC中的签名的潜在临床意义。方法采用实时荧光定量PCR方法检测4种循环血清来源的miRNA（miR-19 a-3 p、miR-203- 3 p、miR-221- 3 p和let-7 f-5 p）在大肠癌患者外周血淋巴结中的表达，并分析其与淋巴结转移的关系。应用受试者工作特征曲线分析其诊断准确性。从人CRC细胞（HCT 116和Caco 2）的条件培养基中分离并表征EV。进行细胞增殖、transwell迁移和管形成测定以研究由CRC-EV转移到内皮细胞中的miR-221- 3 p的促血管生成作用。结果血清来源的miR-19 a-3 p、miR-203- 3 p、miR-221- 3 p和let-7 f-5 p在结直肠癌中的表达水平显著高于健康人。miR-19 a-3 p、miR-203- 3 p和miR-221- 3 p的表达与淋巴结转移状态呈正相关。此外，SOCS 3被鉴定为miR-221- 3 p的直接靶点，CRC-EV穿梭分泌的miR-221- 3 p通过靶向SOCS 3调节内皮细胞中的STAT 3/VEGFR-2信号轴。CRC-EV促进内皮细胞增殖、迁移和血管样结构的形成。CRC-EV对细胞的促血管生成作用通过miR-221- 3 p过表达而重现，显示EV衍生的miR-221- 3 p在促进内皮细胞血管生成中的重要性。结论我们引入了四种循环miRNA（miR-19 a-3 p，miR-203- 3 p，miR-221- 3 p和let-7 f-5 p）作为CRC的新诊断生物标志物。此外，我们发现miR-221- 3 p通过靶向SOCS 3在体外诱导内皮细胞血管生成。
BackgroundSecreted microRNAs (miRNAs) can serve as promising diagnostic markers for colorectal cancer (CRC). Herein, we evaluated the potential clinical significance of a signature of four circulating serum-derived miRNAs in CRC. We also demonstrated that extracellular vesicles (EVs) containing miR-221-3p could facilitate endothelial cell angiogenesis.MethodsThe expressions of four circulating serum-derived miRNAs (miR-19a-3p, miR-203-3p, miR-221-3p, and let-7f-5p) were measured by real-time quantitative PCR, and their associations with lymph node metastasis were determined in CRC patients. Receiver operating characteristic curve analysis was used to determine their diagnostic accuracy. EVs were isolated and characterized from the conditioned media of human CRC cells (HCT116 and Caco2). Cell proliferation, transwell migration, and tube formation assays were performed to investigate the pro-angiogenic effect of miR-221-3p transferred by CRC-EVs into the endothelial cells. In silico analysis was used to show the regulatory functions of miR-221-3p on SOCS3, validated by luciferase and Western blotting assays.ResultsThe expression levels of serum-derived miR-19a-3p, miR-203-3p, miR-221-3p, and let-7f-5p were significantly higher in CRC than in healthy individuals. The expression of miR-19a-3p, miR-203-3p, and miR-221-3p were positively correlated with the lymph node metastasis status. Moreover, SOCS3 was identified as a direct target of miR-221-3p and the secreted miR-221-3p shuttled by CRC-EVs regulated STAT3/VEGFR-2 signaling axis by targeting SOCS3 in endothelial cells. CRC-EVs promoted endothelial cell proliferation, migration, and the formation of vessel-like structures. The proangiogenic effect of CRC-EVs on the cells was recapitulated by miR-221-3p overexpression, showing the importance of EVs-derived miR-221-3p in promoting endothelial cell angiogenesis.ConclusionWe introduced a signature of four-circulating miRNAs (miR-19a-3p, miR-203-3p, miR-221-3p, and let-7f-5p) as a novel diagnostic biomarker for CRC. Besides, we revealed that miR-221-3p induces endothelial cell angiogenesis in vitro by targeting SOCS3.