Eslami-S Zahra, Cortés-Hernández Luis Enrique, Glogovitis Ilias, Antunes-Ferreira Mafalda, D'Ambrosi Silvia, Kurma Keerthi, Garima Françoise, Cayrefourcq Laure, Best Myron G, Koppers-Lalic Danijela, Wurdinger Thomas, Alix-Panabières Catherine
Laboratory of Rare Circulating Human Cells-University Medical Center of Montpellier, Montpellier, France.
CREEC/CANECEV, MIVEGEC (CREES), Université de Montpellier, Centre National de la Recherche Scientifique, Institut de Recherche pour le Développement, Montpellier, France.
Front Cell Dev Biol. 2023 Aug 2;11:1209846. doi: 10.3389/fcell.2023.1209846. eCollection 2023.
Platelets are active players in hemostasis, coagulation and also tumorigenesis. The cross-talk between platelets and circulating tumor cells (CTCs) may have various pro-cancer effects, including promoting tumor growth, epithelial-mesenchymal transition (EMT), metastatic cell survival, adhesion, arrest and also pre-metastatic niche and metastasis formation Interaction with CTCs might alter the platelet transcriptome. However, as CTCs are rare events, the cross-talk between CTCs and platelets is poorly understood. Here, we used our established colon CTC lines to investigate the colon CTC-platelet cross-talk and its impact on the behavior/phenotype of both cell types. We exposed platelets isolated from healthy donors to thrombin (positive control) or to conditioned medium from three CTC lines from one patient with colon cancer and then we monitored the morphological and protein expression changes by microscopy and flow cytometry. We then analyzed the transcriptome by RNA-sequencing of platelets indirectly (presence of a Transwell insert) co-cultured with the three CTC lines. We also quantified by reverse transcription-quantitative PCR the expression of genes related to EMT and cancer development in CTCs after direct co-culture (no Transwell insert) with platelets. We observed morphological and transcriptomic changes in platelets upon exposure to CTC conditioned medium and indirect co-culture (secretome). Moreover, the expression levels of genes involved in EMT ( < 0.05) were decreased in CTCs co-cultured with platelets, but not of genes encoding mesenchymal markers ( and ). The expression levels of genes involved in cancer invasiveness (, and ) were increased. For the first time, we studied the CTC-platelet cross-talk using our unique colon CTC lines. Incubation with CTC conditioned medium led to platelet aggregation and activation, supporting the hypothesis that their interaction may contribute to preserve CTC integrity during their journey in the bloodstream. Moreover, co-culture with platelets influenced the expression of several genes involved in invasiveness and EMT maintenance in CTCs.
血小板在止血、凝血以及肿瘤发生过程中均发挥着积极作用。血小板与循环肿瘤细胞（CTC）之间的相互作用可能具有多种促癌效应，包括促进肿瘤生长、上皮-间质转化（EMT）、转移细胞存活、黏附、停滞，以及促进转移前生态位和转移形成。与CTC的相互作用可能会改变血小板转录组。然而，由于CTC是罕见事件，因此对CTC与血小板之间的相互作用了解甚少。在此，我们利用已建立的结肠CTC系来研究结肠CTC与血小板之间的相互作用及其对两种细胞类型行为/表型的影响。我们将从健康供体分离出的血小板暴露于凝血酶（阳性对照）或来自一名结肠癌患者的三个CTC系的条件培养基中，然后通过显微镜和流式细胞术监测形态和蛋白质表达变化。然后，我们通过对与三个CTC系间接共培养（存在Transwell小室）的血小板进行RNA测序来分析转录组。我们还通过逆转录定量PCR对与血小板直接共培养（无Transwell小室）后CTC中与EMT和癌症发展相关基因的表达进行了定量。我们观察到，血小板暴露于CTC条件培养基和间接共培养（分泌组）后出现了形态和转录组变化。此外，与血小板共培养的CTC中，参与EMT的基因表达水平降低（<0.05），但间充质标志物编码基因（和）的表达水平未降低。参与癌症侵袭的基因（、和）的表达水平升高。我们首次利用我们独特的结肠CTC系研究了CTC与血小板之间的相互作用。与CTC条件培养基孵育导致血小板聚集和激活，支持了它们之间的相互作用可能有助于在血液循环过程中维持CTC完整性的假说。此外，与血小板共培养影响了CTC中参与侵袭和EMT维持的多个基因的表达。
