睾丸生殖细胞肿瘤细胞释放含有microRNA的细胞外囊泡，这些囊泡在肿瘤微环境中诱发表型和基因型变化的细胞。
Testicular germ cell tumour cells release microRNA-containing extracellular vesicles that induce phenotypic and genotypic changes in cells of the tumour microenvironment.
发表日期：2023 Aug 26
作者：
Luz Alonso-Crisostomo, Jennifer Trendell, Marta Ferraresso, Shivani Bailey, Dawn Ward, Zachary G L Scurlock, Stephanie C Wenlock, Carlos A P Bastos, Ravin Jugdaohsingh, Nuno J Faria, Anton J Enright, Cinzia G Scarpini, Nicholas Coleman, Matthew J Murray
来源：
Cellular & Molecular Immunology
摘要：
恶性生殖细胞瘤（GCT）的特征是microRNA（miRNA/miR-）的失调，miR-371~373和miR-302/367簇的过表达是普遍的，不受患者年龄、肿瘤部位或亚型（精原细胞瘤/卵黄囊瘤/胚胎癌）的影响。这些miRNA通过血液释放，假设这是在细胞外小囊泡（EVs）内进行的，并代表了有前景的生物标志物。在这里，我们全面研究了TGCT（睾丸GCT）肿瘤微环境（TME）的（成纤维细胞/内皮/巨噬细胞）细胞与其miRNA载体的EVs间的作用。共进行了34个样本的小RNA下一代测序研究，包括代表性的恶性GCT细胞系/EVs和对照组（睾丸成纤维细胞[Hs1.Tes]细胞系/EVs和睾丸/卵巢样本）。TME细胞接受了TGCT共培养、TGCT源EVs以及miRNA过表达系统（miR-371a-OE）以评估其功能重要性。TGCT细胞将EVs分泌到培养基中。与对照组细胞相比，miR-371~373和miR-302/367簇的miRNA在所有TGCT细胞/亚型中都过表达，并且在TGCT源EVs中丰富，miR-371a-3p/miR-371a-5p最为丰富。TGCT共培养导致TME（成纤维细胞）细胞中miR-371~373和miR-302/367簇miRNA水平增加。接下来，荧光标记证明TGCT源EVs被TME（成纤维细胞/内皮/巨噬细胞）细胞内化。TME（成纤维细胞/内皮）细胞用来自不同TGCT亚型的EVs处理导致miR-371~373和miR-302/367 miRNA水平增加，以及其他通用的（例如，miR-205-5p/miR-148-3p）和亚型特异性的（例如，精原细胞瘤，miR-203a-3p；卵黄囊瘤，miR-375-3p）miRNA。TME细胞中的miR-371a-OE导致胶原蛋白收缩（成纤维细胞）和血管生成（内皮细胞）的增加，通过直接mRNA下调和相关途径的改变。TGCT细胞通过释放富含致癌miRNA的EVs与非肿瘤基质TME细胞进行通信，可能有助于肿瘤进展。© 2023 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.
Malignant germ-cell-tumours (GCTs) are characterised by microRNA (miRNA/miR-) dysregulation, with universal over-expression of miR-371~373 and miR-302/367 clusters regardless of patient age, tumour site, or subtype (seminoma/yolk-sac-tumour/embryonal carcinoma). These miRNAs are released into the bloodstream, presumed within extracellular-vesicles (EVs) and represent promising biomarkers. Here, we comprehensively examined the role of EVs, and their miRNA cargo, on (fibroblast/endothelial/macrophage) cells representative of the testicular GCT (TGCT) tumour microenvironment (TME). Small RNA next-generation-sequencing was performed on 34 samples, comprising representative malignant GCT cell lines/EVs and controls (testis fibroblast [Hs1.Tes] cell-line/EVs and testis/ovary samples). TME cells received TGCT co-culture, TGCT-derived EVs, and a miRNA overexpression system (miR-371a-OE) to assess functional relevance. TGCT cells secreted EVs into culture media. MiR-371~373 and miR-302/367 cluster miRNAs were overexpressed in all TGCT cells/subtypes compared with control cells and were highly abundant in TGCT-derived EVs, with miR-371a-3p/miR-371a-5p the most abundant. TGCT co-culture resulted in increased levels of miRNAs from the miR-371~373 and miR-302/367 clusters in TME (fibroblast) cells. Next, fluorescent labelling demonstrated TGCT-derived EVs were internalised by all TME (fibroblast/endothelial/macrophage) cells. TME (fibroblast/endothelial) cell treatment with EVs derived from different TGCT subtypes resulted in increased miR-371~373 and miR-302/367 miRNA levels, and other generic (eg, miR-205-5p/miR-148-3p) and subtype-specific (seminoma, eg, miR-203a-3p; yolk-sac-tumour, eg, miR-375-3p) miRNAs. MiR-371a-OE in TME cells resulted in increased collagen contraction (fibroblasts) and angiogenesis (endothelial cells), via direct mRNA downregulation and alteration of relevant pathways. TGCT cells communicate with nontumour stromal TME cells through release of EVs enriched in oncogenic miRNAs, potentially contributing to tumour progression.© 2023 The Authors. International Journal of Cancer published by John Wiley & Sons Ltd on behalf of UICC.