Samoto Masahiro, Matsuyama Hideyasu, Matsumoto Hiroaki, Hirata Hiroshi, Ueno Koji, Ozawa Sho, Mori Junichi, Inoue Ryo, Yano Seiji, Yamamoto Yoshiaki, Haginaka Jun, Horiyama Shizuyo, Tamada Koji
Department of Urology, Graduate School of Medicine, Yamaguchi University, Ube, Yamaguchi 755-8505, Japan.
Center for Regenerative Medicine, Graduate School of Medicine, Yamaguchi University, Ube, Yamaguchi 755-8505, Japan.
Oncol Lett. 2021 Oct;22(4):689. doi: 10.3892/ol.2021.12950. Epub 2021 Aug 1.
The interaction between prostate cancer cells and osteoblasts is essential for the development of bone metastasis. Previously, novel androgen receptor axis-targeted agents (ARATs) were approved for metastatic castration-naïve and non-metastatic castration-resistant prostate cancer (CRPC); both of which are pivotal for investigating the association between the bone microenvironment and tumors. The present study established a novel 3D microenvironment model that simulated the bone microenvironment of CRPC, and evaluated the drug susceptibility of ARATs and the efficacy of the combination of abiraterone and dutasteride. Green fluorescent protein-transferred C4-2 cells (a CRPC cell line) and red fluorescent protein-transferred human osteoblasts differentiated from human mesenchymal stem cells were co-cultured in chitosan nanofiber matrix-coated culture plates to simulate the 3D scaffold of the bone microenvironment. The growth of C4-2 was quantified using live-cell imaging and the Cell3 iMager duos analysis system. The growth of C4-2 colonies were quantified for a maximum of 30 days. The expression of TGF-β increased and promoted EMT in C4-2 cells co-cultured with osteoblasts, indicating resistance to ARATs. The IC of each drug and the combination effect of abiraterone and dutasteride were evaluated using this model. Combination treatment with abiraterone and dutasteride synergistically inhibited the growth of C2-4 colonies compared with individual investigational agents. This could be attributed to the reduction of 3-keto-5α-abiraterone, an androgen receptor agonist. The bone microenvironment model of the present study is unique and useful for evaluating new drug susceptibility testing in prostate cancer cells. This model may help to reveal the unknown mechanisms underlying micro- to clinical bone metastasis in prostate cancer.
前列腺癌细胞与成骨细胞之间的相互作用对于骨转移的发生至关重要。此前，新型雄激素受体轴靶向药物（ARATs）已被批准用于转移性去势敏感性和非转移性去势抵抗性前列腺癌（CRPC）；这两者对于研究骨微环境与肿瘤之间的关联都至关重要。本研究建立了一种新型的三维微环境模型，该模型模拟了CRPC的骨微环境，并评估了ARATs的药物敏感性以及阿比特龙和度他雄胺联合用药的疗效。将绿色荧光蛋白转染的C4-2细胞（一种CRPC细胞系）和从人间充质干细胞分化而来的红色荧光蛋白转染的人成骨细胞在壳聚糖纳米纤维基质包被的培养板中共同培养，以模拟骨微环境的三维支架。使用活细胞成像和Cell3 iMager duos分析系统对C4-2细胞的生长进行定量。对C4-2细胞集落的生长进行最长30天的定量。与成骨细胞共同培养的C4-2细胞中转化生长因子-β（TGF-β）的表达增加并促进上皮-间质转化（EMT），表明对ARATs具有抗性。使用该模型评估每种药物的半数抑制浓度（IC）以及阿比特龙和度他雄胺的联合效应。与单独使用研究药物相比，阿比特龙和度他雄胺联合治疗协同抑制了C2-4细胞集落的生长。这可能归因于雄激素受体激动剂3-酮-5α-阿比特龙的减少。本研究的骨微环境模型独特且有助于评估前列腺癌细胞中的新药敏感性测试。该模型可能有助于揭示前列腺癌从微观到临床骨转移的未知机制。
