Establishment and characterization of BHD-F59RSVT, an immortalized cell line derived from a renal cell carcinoma in a patient with Birt-Hogg-Dube syndrome
Establishment and characterization of BHD-F59RSVT, an immortalized cell line derived from a renal cell carcinoma in a patient with Birt-Hogg-Dube syndrome
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DOI：
10.1038/labinvest.2016.137
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发表时间：
2017-03-01
影响因子：
5
通讯作者：
Yao, Masahiro
中科院分区：
文献类型：
--
作者：
Furuya, Mitsuko;Hasumi, Hisashi;Yao, Masahiro
遗传性肾细胞癌(RCC)不仅对患者，而且对他们的亲属都是危及生命的疾病。Birt Hogg-Dube综合征(BHD)是一种由卵泡蛋白基因(Flcn)胚系突变引起的常染色体显性遗传病。这种名为Flcn的蛋白质产物具有肿瘤抑制作用，受影响的患者发展为多个RCC的风险很高。利用啮齿动物模型和人肾细胞癌组织研究了FlcN功能障碍引起的致癌机制。然而，关于基因突变的人肾细胞的体外信号转导的信息非常有限。在此，我们用SV40大T抗原从一例BHD患者的嫌色肾细胞中建立了新的细胞系BHD-F59RSVT。我们研究了该细胞系的FLCN突变、染色体图谱和细胞病理学特征。BHD-F59RSVT反映了患者的FLCN胚系突变，即外显子13(c.1528_1530delGAG)的3个碱基缺失。未检测到FLCN的体细胞突变和杂合性丢失。FLCN近端的17p11.2号染色体呈三峰模式。全基因组染色体分析显示BHD-F59RSVT细胞16号染色体缺失，7号染色体出现嵌合性片段。BHD-F59RSVT细胞角蛋白7免疫组化染色阳性，支持其起源于远端曲管。Western blotting分析表明，Flcn在蛋白水平上受到严重抑制。这些集体发现表明，所建立的细胞系将适合于BHD相关肾癌典型表型的功能分析，该表型抑制了Flcn的表达。
Hereditary renal cell carcinomas (RCCs) are life-threatening disorders not only for the patients but also for their relatives. Birt Hogg-Dube syndrome (BHD) is an autosomal dominant disorder caused by germline mutations in the folliculin gene (FLCN). The protein product, FLCN, functions as a tumor suppressor, and the affected patients have high risks of developing multiple RCCs. The carcinogenic mechanisms stemming from FLCN dysfunction have been investigated using rodent models and human RCC tissues. However, very limited information has been available about in vitro signaling of human renal cells with genetically mutant FLCN. Herein, we established a new cell line, BHD-F59RSVT, from a BHD patient's chromophobe RCC by transfecting SV40 large T antigen. We investigated FLCN mutations, chromosome profiles, and cytopathologic characteristics of the cell line. BHD-F59RSVT reflected the patient's FLCN germline mutation, a 3-nt deletion in exon 13 (c.1528_1530delGAG). Neither somatic mutation nor loss of heterozygosity of FLCN was detectable. Chromosome 17p11.2 of the FLCN proximal region demonstrated a trimodal pattern. Genome-wide chromosomal analysis revealed a loss of chromosome 16 and mosaic segmental gains in chromosome 7. BHD-F59RSVT cells were positive when immunostained for cytokeratin 7, supporting their origin from distal convoluted tubules. Western blotting analysis demonstrated severely suppressed FLCN expression at the protein level. The collective findings indicate that the established cell line will be suitable for functional analysis of the typical phenotype of BHD-associated RCC with suppressed FLCN expression.