EMT-cancer cells-derived exosomal miR-27b-3p promotes circulating tumour cells-mediated metastasis by modulating vascular permeability in colorectal cancer.
EMT-cancer cells-derived exosomal miR-27b-3p promotes circulating tumour cells-mediated metastasis by modulating vascular permeability in colorectal cancer.
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DOI：
10.1002/ctm2.595
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发表时间：
2021-12
影响因子：
10.6
通讯作者：
Xiong B
中科院分区：
文献类型：
--
作者：
Dou R;Liu K;Yang C;Zheng J;Shi D;Lin X;Wei C;Zhang C;Fang Y;Huang S;Song J;Wang S;Xiong B
转移是结直肠癌(CRC)的主要死亡原因。循环肿瘤细胞(CTCs)被认为是肿瘤转移的前体细胞。CTCs经历了上皮-间充质转化(EMT)，与转移有关，预后不良。EMT癌细胞调节侵袭前沿内皮细胞的通透性，促进癌细胞内皮细胞的侵入，导致CTCs介导的远处转移。来源于癌细胞的外切体是肿瘤-宿主相互交流的关键媒介。然而，EMT-肿瘤细胞来源的外切体调节血管通透性和促进CTCs生成的机制尚不清楚。采用超速离心法分离纯化外切体。通过测序和定量聚合酶链式反应对外体miRNA进行鉴定。体外共培养实验观察外体miR-27b-3p对血管内皮细胞通透性的影响。采用双荧光素酶报告实验、染色质免疫沉淀(ChIP)和RNA免疫沉淀(RIP)等方法研究miR-27b-3p被包装到外体中的可能机制。建立小鼠模型，研究外体miR-27b-3p在体内血管通透性调节中的作用。我们发现EMT-CRC细胞通过将miR-27b-3p转移到外体中的人脐静脉内皮细胞(HUVECs)来减弱血管屏障。从机制上讲，miR-27b-3p通过直接与VE-Cad和p120的3‘非翻译区结合，在转录后水平抑制VE-Cad和p120的表达。MiR-27b-3p被STAT3激活的异质核核糖核蛋白A1(HnRNPA1)诱导包装到外体中。临床上，miR-27b-3p在大肠癌组织中表达上调。血浆外体miR-27b-3p与结直肠癌恶性进展及CTC计数呈正相关。我们的研究揭示了EMT-CRC细胞促进转移、增加血管通透性和促进CTCs生成的新机制。EMT-CRC细胞分泌的外体miR-27b-3p可增加血管通透性，促进CTCs的生成。胞外体miR-27b-3p有望成为判断结直肠癌转移的生物标志物。综上所述，我们的研究结果显示，EMT CRC细胞来源的外体miR-27b-3p转移到内皮细胞，增强血管通透性，从而通过靶向VE-cadherin和p120促进CTC的生成。上游STAT3通过结合其启动子上调hnRNPA1的表达，hnRNPA1介导miR-27b-3p的包装。我们的研究强调，STAT3/hnRNPA1/miR-27b-3p信号级联和外体miR-27b-3p可能是诊断结直肠癌转移的有前景的生物标志物。
Metastasis is the main cause of death in colorectal cancer (CRC). Circulating tumour cells (CTCs) are regarded as the precursor cells of metastasis. The CTCs, which underwent epithelial‐mesenchymal transition (EMT), are associated with metastasis and responsible for poor prognosis. EMT cancer cells modulate endothelial permeability in the invasive front and facilitate cancer cell intravasation, resulting in CTCs‐mediated distant metastasis. Exosomes derived from cancer cells are key mediators of cancer‐host intercommunication. However, the mechanism by which EMT‐tumour cells‐derived exosomes modulate vascular permeability and promote CTCs generation has remained unclear. Exosomes isolation and purification were conducted by ultra‐centrifugation. Exosomal miRNA was identified by sequencing followed by quantitative PCR. In vitro co‐culture assay experiments were conducted to evaluate the effect of exosomal miR‐27b‐3p on the permeability of blood vessel endothelium. Dual‐luciferase reporter assay, chromatin immunoprecipitation (ChIP) and RNA immunoprecipitation (RIP) were performed to investigate the underlying mechanism by which miR‐27b‐3p is packaged into exosomes. A mouse model was established to determine the role of exosomal miR‐27b‐3p in blood vessel permeability modulation in vivo. We found that EMT‐CRC cells attenuate the blood vessel barrier by transferring miR‐27b‐3p to human umbilical vein endothelial cells (HUVECs) in exosomes. Mechanically, miR‐27b‐3p atteuated the expression of vascular endothelial cadherin (VE‐Cad) and p120 at the post‐transcriptional level by binding to 3′‐untranslated region of VE‐Cad and p120 directly. The packaging of miR‐27b‐3p into exosomes was induced by heterogeneous nuclear ribonucleoprotein A1 (hnRNPA1), which activated by STAT3. Clinically, miR‐27b‐3p up‐regulated in CRC tissues. Plasma exosomal miR‐27b‐3p was positively correlated with malignant progression and CTC count in CRC patients. Our study reveals a novel mechanism by which EMT‐CRC cells promote metastasis, increasing blood vessel permeability and facilitating the generation of CTCs. Exosomal miR‐27b‐3p secreted by EMT‐CRC cells increases blood vessel permeability and facilitates the generation of CTCs. Exosomal miR‐27b‐3p may become a promising biomarker for CRC metastasis. Collectively, our findings revealed that EMT CRC cell‐derived exosomal miR‐27b‐3p transferred to endothelial cells, enhanced blood vessel permeability, and thus facilitated CTC generation by targeting VE‐cadherin and p120. Upstream STAT3 upregulated hnRNPA1 by binding its promotor and hnRNPA1 mediated the packaging of miR‐27b‐3p. Our study highlighted that the STAT3/hnRNPA1/miR‐27b‐3p signal cascade and exosomal miR‐27b‐3p may be promising diagnostic biomarkers for CRC metastasis.
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影响因子：
4
作者：
Dejana, Elisabetta;Orsenigo, Fabrizio;Lampugnani, Maria Grazia
通讯作者：
Lampugnani, Maria Grazia
DOI：
10.1073/pnas.1524448113
发表时间：
2016-05-03
影响因子：
11.1
作者：
Au, Sam H.;Storey, Brian D.;Toner, Mehmet
通讯作者：
Toner, Mehmet
DOI：
10.1158/1078-0432.ccr-13-3173
发表时间：
2015-03-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Jung HY;Fattet L;Yang J
通讯作者：
Yang J
影响因子：
11.2
作者：
Caetano MS;Zhang H;Cumpian AM;Gong L;Unver N;Ostrin EJ;Daliri S;Chang SH;Ochoa CE;Hanash S;Behrens C;Wistuba II;Sternberg C;Kadara H;Ferreira CG;Watowich SS;Moghaddam SJ
通讯作者：
Moghaddam SJ
影响因子：
8.8
作者：
Koppers-Lalic, Danijela;Hackenberg, Michael;Pegtel, D. Michiel
通讯作者：
Pegtel, D. Michiel