Feely Sarah, Mullen Nathan, Donlon Padraig T, Reidy Eileen, Challapalli Ritihaas Surya, Hassany Mariam, Sorushanova Anna, Martinez Eduardo Ribes, Owens Peter, Quinn Anne Marie, Pandit Abhay, Harhen Brendan, Finn David P, Hantel Constanze, O'Halloran Martin, Prakash Punit, Dennedy Michael C
Discipline of Pharmacology and Therapeutics, School of Medicine, University of Galway, Galway, H91 V4AY, Ireland.
Science Foundation Ireland (SFI) Research Centre for Research in Medical Devices (CURAM), Biomedical Science Building, University of Galway, Galway, H91 TK33, Ireland.
Endocrinology. 2024 Nov 26;166(1). doi: 10.1210/endocr/bqae159.
Adrenocortical carcinoma (ACC) is a rare malignancy of the adrenal cortex that is associated with a poor prognosis. Developing effective treatment options for ACC is challenging owing to the current lack of representative preclinical models. This study addressed this limitation by developing and characterizing 3-dimensional (3D) cell cultures incorporating the ACC cell lines, MUC-1, HAC15, and H295R in a type I collagen matrix. ACC tissue samples were analyzed by immunohistochemistry to determine the presence of type I collagen in the tumor microenvironment. Cell viability and proliferation were assessed using flow cytometry and confocal microscopy. mRNA expression of steroidogenic enzymes and steroid secretion was analyzed by comparing the 3D and monolayer cell culture models. All cells were successfully cultured in a type I collagen matrix, which is highly expressed in the ACC tumor microenvironment and showed optimal viability until day 7. All 3 models showed increased metabolic and proliferative activity over time. Three-dimensional cell cultures were steroidogenic and demonstrated increased resistance to the gold standard chemotherapy, mitotane, compared with monolayer. The use of these models may lead to an improved understanding of disease pathology and provide a better representative platform for testing and screening of potential therapies.
肾上腺皮质癌（ACC）是一种罕见的肾上腺皮质恶性肿瘤，预后较差。由于目前缺乏具有代表性的临床前模型，开发针对ACC的有效治疗方案具有挑战性。本研究通过在I型胶原基质中培养并鉴定包含ACC细胞系MUC-1、HAC15和H295R的三维（3D）细胞培养物，解决了这一局限性。通过免疫组织化学分析ACC组织样本，以确定肿瘤微环境中I型胶原的存在。使用流式细胞术和共聚焦显微镜评估细胞活力和增殖。通过比较3D和单层细胞培养模型，分析类固醇生成酶的mRNA表达和类固醇分泌。所有细胞均在I型胶原基质中成功培养，该基质在ACC肿瘤微环境中高度表达，并在第7天前显示出最佳活力。所有3种模型均显示随着时间推移代谢和增殖活性增加。与单层细胞相比，三维细胞培养具有类固醇生成能力，并且对金标准化疗药物米托坦表现出更高的耐药性。使用这些模型可能会增进对疾病病理学的理解，并为测试和筛选潜在疗法提供更好的代表性平台。
