Piquet Léo, Dewit Louise, Schoonjans Nathan, Millet Martial, Bérubé Julie, Gerges Peter R A, Bordeleau François, Landreville Solange
Faculté de médecine, Université Laval, Quebec City, QC G1V 0A6, Canada.
Centre de recherche du CHU de Québec-Université Laval, Quebec City, QC G1S 4L8, Canada.
Cancers (Basel). 2019 Jul 24;11(8):1043. doi: 10.3390/cancers11081043.
Uveal melanoma (UM) is a malignant intraocular tumor that spreads to the liver in half of the cases. Since hepatic cells could play a role in the therapeutic resistance of metastatic UM, the purpose of our study was to investigate the pro-invasive role of hepatic stellate cells (HSteCs) in metastatic UM at the micro- and macro-metastatic stages. We first performed an immunostaining with the alpha-smooth muscle actin (αSMA) to localize activated HSteCs in UM liver macro-metastases from four patients. Their accumulation of collagen was assessed with Masson's Trichrome stain. Next, we inoculated metastatic UM cells alone or with human HSteCs in triple-immunodeficient mice, in order to determine if HSteCs are recruited as early as the micro-metastatic stage. The growth of metastatic foci was imaged in the liver by fluorescence imaging. Histological analyses were performed with Masson's Trichrome and Picrosirius Red stains, and antibodies against Melan-A and αSMA. The collagen content was measured in xenografts by quantitative polarization microscopy. In patient hepatectomy samples, activated HSteCs and their pathological matrix were localized surrounding the malignant lesions. In the mouse xenograft model, the number of hepatic metastases was increased when human HSteCs were co-inoculated. Histological analyses revealed a significant recruitment of HSteCs near the micro/macrolesions, and an increase in fibrillar collagen production. Our results show that HSteCs can provide a permissive microenvironment and might increase the therapeutic resistance of metastatic UM.
葡萄膜黑色素瘤（UM）是一种恶性眼内肿瘤，半数病例会转移至肝脏。由于肝细胞可能在转移性UM的治疗抵抗中发挥作用，我们研究的目的是在微转移和大转移阶段研究肝星状细胞（HSteCs）在转移性UM中的促侵袭作用。我们首先用α-平滑肌肌动蛋白（αSMA）进行免疫染色，以定位4例患者UM肝大转移灶中活化的HSteCs。用Masson三色染色评估其胶原积累情况。接下来，我们将转移性UM细胞单独或与人HSteCs一起接种到三联免疫缺陷小鼠体内，以确定HSteCs是否早在微转移阶段就被募集。通过荧光成像对肝脏中转移灶的生长进行成像。用Masson三色染色和天狼星红染色以及抗Melan-A和αSMA抗体进行组织学分析。通过定量偏振显微镜测量异种移植瘤中的胶原含量。在患者肝切除样本中，活化的HSteCs及其病理基质定位于恶性病变周围。在小鼠异种移植模型中，与人HSteCs共同接种时肝转移数量增加。组织学分析显示在微/大病变附近有大量HSteCs被募集，并且纤维状胶原产生增加。我们的结果表明，HSteCs可以提供一个有利的微环境，并可能增加转移性UM的治疗抵抗。
