New chondrosarcoma cell lines and mouse models to study the link between chondrogenesis and chemoresistance
New chondrosarcoma cell lines and mouse models to study the link between chondrogenesis and chemoresistance
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DOI：
10.1038/labinvest.2013.101
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发表时间：
2013-10-01
影响因子：
5
通讯作者：
Blanchard, Frederic
中科院分区：
文献类型：
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作者：
Monderer, David;Luseau, Alexandrine;Blanchard, Frederic
软骨肉瘤是一种软骨形成、血管稀少的肿瘤。骨肉瘤是成人仅次于骨肉瘤的第二种恶性原发骨肿瘤，但与骨肉瘤相比，骨肉瘤对化疗和放射治疗耐药，手术切除仍是唯一的治疗选择。几乎没有可用的细胞系和动物模型，其化疗耐药背后的机制在很大程度上仍不清楚。我们的目标是从人软骨肉瘤活检组织中建立新的细胞系和动物癌症模型，以研究它们的化疗耐药性。2007年至2012年，收集了10例软骨肉瘤活检组织，用于细胞培养和移植到裸鼠体内。只有一例移植活检和一株注射细胞系移植成功，导致了与原始活检相似的常规中央型高级别软骨肉瘤。在培养中，获得了两个新的稳定的细胞系，一个来自去分化的软骨肉瘤，另一个来自常规的III级中央软骨肉瘤活检。其基因特征为三倍体核型，IDH1、IDH2和TP53突变，CDKN2A缺失和/或MDM2扩增。这些细胞株表达CD44、73、90、105等细胞膜标志物，在软骨化的三维颗粒中培养可形成透明的软骨基质。使用高通量定量RT-PCR方法，我们观察到单层培养的细胞株失去了几个与软骨发育有关的基因(COL2A1、COMP、ACAN)的表达，但在3D培养中恢复了它们的表达。单层软骨肉瘤细胞对几种常规化疗药物敏感，但在3D颗粒中培养时，随着药物核积累的改变，对低剂量的马福福胺或阿霉素产生抗药性。我们的结果表明，软骨肉瘤细胞产生的软骨基质可能会损害几种药物的扩散，从而导致化疗耐药。因此，3D软骨细胞颗粒是研究软骨肉瘤化疗耐药的一种更具相关性的模型，可能是一种有价值的动物实验替代方法。
Chondrosarcomas are cartilage-forming, poorly vascularized tumors. They represent the second malignant primary bone tumor of adults after osteosarcoma, but in contrast to osteosarcoma they are resistant to chemotherapy and radiotherapy, surgical excision remaining the only therapeutic option. Few cell lines and animal models are available, and the mechanisms behind their chemoresistance remain largely unknown. Our goal was to establish new cell lines and animal cancer models from human chondrosarcoma biopsies to study their chemoresistance. Between 2007 and 2012, 10 chondrosarcoma biopsies were collected and used for cell culture and transplantation into nude mice. Only one transplanted biopsy and one injected cell line has engrafted successfully leading to conventional central high-grade chondrosarcoma similar to the original biopsies. In culture, two new stable cell lines were obtained, one from a dedifferentiated and one from a grade III conventional central chondrosarcoma biopsy. Their genetic characterization revealed triploid karyotypes, mutations in IDH1, IDH2, and TP53, deletion in CDKN2A and/or MDM2 amplification. These cell lines expressed nnesenchynnal membrane markers (CD44, 73, 90, 105) and were able to produce a hyaline cartilaginous matrix when cultured in chondrogenic three-dimensional (3D) pellets. Using a high-throughput quantitative RT-PCR approach, we observed that cell lines cultured in monolayer had lost expression of several genes implicated in cartilage development (COL2A1, COMP, ACAN) but restored their expression in 3D cultures. Chondrosarcoma cells in monolayer were sensitive to several conventional chemotherapeutic agents but became resistant to low doses of mafosfamide or doxorubicin when cultured in 3D pellets, in parallel with an altered nucleic accumulation of the drug. Our results indicate that the cartilaginous matrix produced by chondrosarcoma cells may impair diffusion of several drugs and thus contribute to chemoresistance. Therefore, 3D chondrogenic cell pellets constitute a more relevant model to study chondrosarcoma chemoresistance and may be a valuable alternative to animal experimentations.