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首个携带嵌合型SS18-SSX1基因的克隆人类永生化脂肪来源间充质干细胞系(SS-iASC)。

First cloned human immortalized adipose derived mesenchymal stem-cell line with chimeric SS18-SSX1 gene (SS-iASC).

作者信息

Mihály Dóra, Matula Zsolt, Changchien Yi-Che, Papp Gergő, Tátrai Péter, Sápi Zoltán

机构信息

1st Department of Pathology and Experimental Cancer Research, Semmelweis University, Üllői út 26, H-1085 Budapest, Hungary.

Institute of Enzymology, Research Center for Natural Sciences, Hungarian Academy of Sciences, Magyar tudósok körútja 2, H-1117 Budapest, Hungary.

出版信息

Cancer Genet. 2017 Oct;216-217:52-60. doi: 10.1016/j.cancergen.2017.07.003. Epub 2017 Jul 26.

DOI:10.1016/j.cancergen.2017.07.003
PMID:29025595
Abstract

The SS18-SSX chimeric gene is unique to synovial sarcoma. Multiple model systems including mouse cell lines expressing SS18-SSX, and genetically engineered mouse models of synovial sarcoma have been developed to elucidate the role of the chimeric gene in synovial sarcomagenesis. Although several cell lines stably expressing human SS18-SSX exist, there is an ongoing need for cell culture models enabling researchers to investigate the molecular mechanism of SS18-SSX action in a relevant cellular context. Here we report the establishment of a novel SS18-SSX1-expressing cell line created from immortalized human adipose tissue-derived mesenchymal stem cells via lentiviral transduction of the chimeric gene. Our cell line, termed SS-iASC, has been characterized by karyotyping and cell line identification, and stable expression of SS18-SSX1 has been verified using real-time PCR (RT-PCR), nested PCR, immunofluorescence, and immunoblotting. Focal cytokeratin positivity characteristic of synovial sarcoma but no β-Catenin, Bcl-2 or cyclin D1 expression was observed in SS-iASC. The novel cell line expressing SS18-SSX1 on a human adipose-derived stromal cell background is expected to be helpful in addressing the question whether the chimeric gene alone is sufficient to trigger the formation of synovial sarcoma.

摘要

SS18-SSX嵌合基因是滑膜肉瘤所特有的。已经开发了多种模型系统,包括表达SS18-SSX的小鼠细胞系以及滑膜肉瘤的基因工程小鼠模型,以阐明嵌合基因在滑膜肉瘤发生中的作用。尽管存在几种稳定表达人SS18-SSX的细胞系,但仍持续需要细胞培养模型,以使研究人员能够在相关细胞背景下研究SS18-SSX作用的分子机制。在此,我们报告通过慢病毒转导嵌合基因从永生化的人脂肪组织来源的间充质干细胞创建了一种新型的表达SS18-SSX1的细胞系。我们的细胞系称为SS-iASC,已通过核型分析和细胞系鉴定进行了表征,并使用实时PCR(RT-PCR)、巢式PCR、免疫荧光和免疫印迹验证了SS18-SSX1的稳定表达。在SS-iASC中观察到滑膜肉瘤特有的局灶性细胞角蛋白阳性,但未观察到β-连环蛋白、Bcl-2或细胞周期蛋白D1的表达。预计在人脂肪来源的基质细胞背景上表达SS18-SSX1的新型细胞系将有助于解决仅嵌合基因是否足以引发滑膜肉瘤形成的问题。

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First cloned human immortalized adipose derived mesenchymal stem-cell line with chimeric SS18-SSX1 gene (SS-iASC).首个携带嵌合型SS18-SSX1基因的克隆人类永生化脂肪来源间充质干细胞系(SS-iASC)。
Cancer Genet. 2017 Oct;216-217:52-60. doi: 10.1016/j.cancergen.2017.07.003. Epub 2017 Jul 26.
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Downregulation of SS18-SSX1 expression in synovial sarcoma by small interfering RNA enhances the focal adhesion pathway and inhibits anchorage-independent growth in vitro and tumor growth in vivo.通过小干扰 RNA 下调滑膜肉瘤中 SS18-SSX1 的表达可增强黏着斑通路,抑制体外无锚定生长和体内肿瘤生长。
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Loss of SS18-SSX1 inhibits viability and induces apoptosis in synovial sarcoma.SS18-SSX1 缺失抑制滑膜肉瘤细胞活力并诱导其凋亡。
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SS18-SSX fusion protein-induced Wnt/β-catenin signaling is a therapeutic target in synovial sarcoma.SS18-SSX融合蛋白诱导的Wnt/β-连环蛋白信号传导是滑膜肉瘤的一个治疗靶点。
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