Generation of chordoma cell line JHC7 and the identification of Brachyury as a novel molecular target.
Generation of chordoma cell line JHC7 and the identification of Brachyury as a novel molecular target.
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DOI：
10.3171/2011.5.jns11185
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发表时间：
2011-10
影响因子：
4.1
通讯作者：
Quiñones-Hinojosa A
中科院分区：
文献类型：
--
作者：
Hsu W;Mohyeldin A;Shah SR;ap Rhys CM;Johnson LF;Sedora-Roman NI;Kosztowski TA;Awad OA;McCarthy EF;Loeb DM;Wolinsky JP;Gokaslan ZL;Quiñones-Hinojosa A
脊索瘤是一种恶性骨肿瘤，被认为是沿着神经轴的脊索残留物。最近对脊索瘤的基因组研究已经确定T（Brachyury）基因重复是家族性脊索瘤的主要易感突变。Brachyury在脊索的胚胎发育过程中起着至关重要的作用，最近已被证明可以调节上皮来源的癌症中的上皮-间充质转化。然而，由于缺乏表达Brachyury的完全表征的脉络膜细胞系，目前对该转录因子在脉络膜中的作用的理解是有限的。因此，本研究的目的是建立第一个完全表征的原代脉络膜细胞系，其表达T基因位点的增益，其容易在体外和体内重现原始亲本肿瘤表型。使用来自患有原发性骶骨脉络膜炎的61岁女性的术中获得的肿瘤样品，建立了脉络膜细胞系（JHC 7，或约翰霍普金斯脉络膜系7）。使用标准免疫染色和蛋白质印迹进行原发性肿瘤和细胞系的分子表征。测定了该细胞系中的染色体畸变和T基因的基因组扩增。使用该细胞系，建立异种移植模型，并进行肿瘤的组织病理学分析。评估Brachyury的沉默和基因表达的变化。作者报告，首次成功建立了一个脉络膜细胞系（JHC 7），从病理证实的骶骨脉络膜。该细胞系容易在免疫缺陷小鼠中形成肿瘤，其重现了具有与亲本肿瘤一致的保守组织学特征的亲本肿瘤表型。此外，首次证明了使用短发夹RNA沉默Brachyury使脉络膜细胞的形态呈现更分化的样状态，并导致完全生长停滞和衰老，无法在体外连续传代。本报告代表了文献中描述的第一个骶骨脉络膜细胞系的异种移植模型和第一个建立的具有稳定Brachyury表达的细胞系。作者提出，Brachyury是一个有吸引力的治疗目标，在脉络膜和JHC7将作为一个临床相关的模型，这种疾病的研究。
Chordoma is a malignant bone neoplasm hypothesized to arise from notochordal remnants along the length of the neuraxis. Recent genomic investigation of chordomas has identified T (Brachyury) gene duplication as a major susceptibility mutation in familial chordomas. Brachyury plays a vital role during embryonic development of the notochord and has recently been shown to regulate epithelial-to-mesenchymal transition in epithelial-derived cancers. However, current understanding of the role of this transcription factor in chordoma is limited due to the lack of availability of a fully characterized chordoma cell line expressing Brachyury. Thus, the objective of this study was to establish the first fully characterized primary chordoma cell line expressing gain of the T gene locus that readily recapitulates the original parental tumor phenotype in vitro and in vivo. Using an intraoperatively obtained tumor sample from a 61-year-old woman with primary sacral chordoma, a chordoma cell line (JHC7, or Johns Hopkins Chordoma Line 7) was established. Molecular characterization of the primary tumor and cell line was conducted using standard immunostaining and Western blotting. Chromosomal aberrations and genomic amplification of the T gene in this cell line were determined. Using this cell line, a xenograft model was established and the histopathological analysis of the tumor was performed. Silencing of Brachyury and changes in gene expression were assessed. The authors report, for the first time, the successful establishment of a chordoma cell line (JHC7) from a patient with pathologically confirmed sacral chordoma. This cell line readily forms tumors in immunodeficient mice that recapitulate the parental tumor phenotype with conserved histological features consistent with the parental tumor. Furthermore, it is demonstrated for the first time that silencing of Brachyury using short hairpin RNA renders the morphology of chordoma cells to a more differentiated-like state and leads to complete growth arrest and senescence with an inability to be passaged serially in vitro. This report represents the first xenograft model of a sacral chordoma line described in the literature and the first cell line established with stable Brachyury expression. The authors propose that Brachyury is an attractive therapeutic target in chordoma and that JHC7 will serve as a clinically relevant model for the study of this disease.