title: Establishment and characterization of BHD-F59RSVT, an immortalized cell line derived from a renal cell carcinoma in a patient with Birt-Hogg-Dubé syndrome.
abstract: Hereditary renal cell carcinomas (RCCs) are life-threatening disorders not only for the patients but also for their relatives. Birt-Hogg-Dubé 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.
authors.affiliation: Department of Molecular Pathology, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
authors.forename: Mitsuko
authors.initials: M
authors.lastname: Furuya
authors.affiliation: Department of Urology, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
authors.forename: Hisashi
authors.initials: H
authors.lastname: Hasumi
authors.affiliation: International Research Center for Medical Sciences (IRCMS), Kumamoto University, Kumamoto, Japan.
authors.forename: Masaya
authors.initials: M
authors.lastname: Baba
authors.affiliation: Medical Mycology Research Center, Chiba University, Chiba, Japan.
authors.forename: Reiko
authors.initials: R
authors.lastname: Tanaka
authors.affiliation: Department of Molecular Pathology, Yokohama City University Graduate School of Medicine, Yokohama, Japan.|Department of Urology, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Iribe
authors.affiliation: Department of Internal Medicine, Ise Red Cross Hospital, Ise, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Onishi
authors.affiliation: Department of Surgical Pathology, Tokyo Women's Medical University, Tokyo, Japan.
authors.forename: Yoji
authors.initials: Y
authors.lastname: Nagashima
authors.affiliation: Department of Diagnostic Pathology, Chiba University Graduate School of Medicine, Chiba, Japan.
authors.forename: Yukio
authors.initials: Y
authors.lastname: Nakatani
authors.affiliation: Department of Otolaryngology, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
authors.forename: Yasuhiro
authors.initials: Y
authors.lastname: Isono
authors.affiliation: Department of Urology, Yokohama City University Graduate School of Medicine, Yokohama, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yao
mesh_terms: D058249:Birt-Hogg-Dube Syndrome / Q000150:complications / Q000235:genetics*; D002292:Carcinoma, Renal Cell / Q000150:complications / Q000235:genetics* / Q000473:pathology; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D056915:DNA Copy Number Variations; D004252:DNA Mutational Analysis / Q000379:methods; D005192:Family Health; D005260:Female; D018095:Germ-Line Mutation; D006801:Humans; D007680:Kidney Neoplasms / Q000150:complications / Q000235:genetics* / Q000473:pathology; D019656:Loss of Heterozygosity; D008297:Male; D008875:Middle Aged; D010375:Pedigree; D011518:Proto-Oncogene Proteins / Q000235:genetics* / Q000378:metabolism; D032681:Spectral Karyotyping / Q000379:methods; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: C476078:FLCN protein, human; D011518:Proto-Oncogene Proteins; D025521:Tumor Suppressor Proteins