title: Fumarate Hydratase-deficient Cell Line NCCFH1 as a New In Vitro Model of Hereditary Papillary Renal Cell Carcinoma Type 2.
abstract: OBJECTIVE
  Hereditary leiomyomatosis and renal cell carcinoma (HLRCC) is a rare autosomal dominant disorder characterized by fumarate hydratase (FH) gene mutation. It is associated with the development of very aggressive kidney tumors, characterized by early onset and high metastatic potential, and has no effective therapy. The aim of the study was to establish a new preclinical platform for investigating morphogenetic and metabolic features, and alternative therapy of metastatic hereditary papillary renal cell carcinoma type 2 (PRCC2).
  
  
  METHODS
  Fresh cells were collected from pleural fluid of a patient with metastatic hereditary PRCC2. Morphogenetic and functional characteristics were evaluated via microscopy, FH gene sequencing analysis, real-time polymerase chaine reaction and enzymatic activity measurement. We performed bioenergetic analysis, gene-expression profiling, and cell viability assay with 19 anti-neoplastic drugs.
  
  
  RESULTS
  We established a new in vitro model of hereditary PRCC2 - the NCCFH1 cell line. The cell line possesses a c.1162 delA - p.Thr375fs frameshift mutation in the FH gene. Our findings indicate severe attenuation of oxidative phosphorylation and glucose-dependent growth of NCCFH1 cells that is consistent with the Warburg effect. Furthermore, gene-expression profiling identified that the most prominent molecular features reflected a high level of apoptosis, cell adhesion, and cell signaling. Drug screening revealed a marked sensitivity of FH(-/-) cells to mitoxantrone, epirubicin, topotecan and a high sensitivity to bortezomib.
  
  
  CONCLUSIONS
  We demonstrated that the NCCFH1 cell line is a very interesting preclinical model for studying the metabolic features and testing new therapies for hereditary PRCC2, while bortezomib may be a potential efficient therapeutic option.
authors.affiliation: Ecole Pratique des Hautes Etudes, Paris and French National Institute of Health and Medical Research U753, Gustave Roussy Cancer Campus, Villejuif, France Division of Medical Sciences, National Cancer Centre Singapore, Singapore, Republic of Singapore.
authors.forename: Victoria
authors.initials: V
authors.lastname: Perrier-Trudova
authors.affiliation: Division of Medical Sciences, National Cancer Centre Singapore, Singapore, Republic of Singapore Division of Cancer and Stem Cell Biology, Duke-NUS Graduate Medical School, Singapore, Republic of Singapore.
authors.forename: Bernice Wong
authors.initials: BW
authors.lastname: Huimin
authors.affiliation: Division of Medical Sciences, National Cancer Centre Singapore, Singapore, Republic of Singapore.
authors.forename: Sarinya
authors.initials: S
authors.lastname: Kongpetch
authors.affiliation: Division of Medical Sciences, National Cancer Centre Singapore, Singapore, Republic of Singapore.
authors.forename: Dachuan
authors.initials: D
authors.lastname: Huang
authors.affiliation: Division of Medical Sciences, National Cancer Centre Singapore, Singapore, Republic of Singapore Division of Cancer and Stem Cell Biology, Duke-NUS Graduate Medical School, Singapore, Republic of Singapore.
authors.forename: Pauline
authors.initials: P
authors.lastname: Ong
authors.affiliation: Ecole Pratique des Hautes Etudes, Paris and French National Institute of Health and Medical Research U753, Gustave Roussy Cancer Campus, Villejuif, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Le Formal
authors.affiliation: Division of Medical Sciences, National Cancer Centre Singapore, Singapore, Republic of Singapore Division of Cancer and Stem Cell Biology, Duke-NUS Graduate Medical School, Singapore, Republic of Singapore.
authors.forename: Song Ling
authors.initials: SL
authors.lastname: Poon
authors.affiliation: Division of Medical Sciences, National Cancer Centre Singapore, Singapore, Republic of Singapore.
authors.forename: Ee Yan
authors.initials: EY
authors.lastname: Siew
authors.affiliation: Division of Medical Sciences, National Cancer Centre Singapore, Singapore, Republic of Singapore.
authors.forename: Swe Swe
authors.initials: SS
authors.lastname: Myint
authors.affiliation: Ecole Pratique des Hautes Etudes, Paris and French National Institute of Health and Medical Research U753, Gustave Roussy Cancer Campus, Villejuif, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Gad
authors.affiliation: French National Institute of Health and Medical Research U892 and French National Centre for Scientific Research 6299, Cancer Research Center Nantes-Angers, University of Nantes, Nantes, France.
authors.forename: Betty
authors.initials: B
authors.lastname: Gardie
authors.affiliation: Ecole Pratique des Hautes Etudes, Paris and French National Institute of Health and Medical Research U753, Gustave Roussy Cancer Campus, Villejuif, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Couvé
authors.affiliation: Department of Biodevices and Diagnostics, Institute of Bioengineering and Nanotechnology, Singapore, Republic of Singapore.
authors.forename: Yu Miin
authors.initials: YM
authors.lastname: Foong
authors.affiliation: Department of Biodevices and Diagnostics, Institute of Bioengineering and Nanotechnology, Singapore, Republic of Singapore.
authors.forename: Yukti
authors.initials: Y
authors.lastname: Choudhury
authors.affiliation: Department of Biodevices and Diagnostics, Institute of Bioengineering and Nanotechnology, Singapore, Republic of Singapore.
authors.forename: Jonathan
authors.initials: J
authors.lastname: Poh
authors.affiliation: Division of Medical Sciences, National Cancer Centre Singapore, Singapore, Republic of Singapore Division of Cancer and Stem Cell Biology, Duke-NUS Graduate Medical School, Singapore, Republic of Singapore.
authors.forename: Choon Kiat
authors.initials: CK
authors.lastname: Ong
authors.affiliation: Division of Medical Oncology, National Cancer Centre Singapore, Singapore, Republic of Singapore.
authors.forename: Chee Keong
authors.initials: CK
authors.lastname: Toh
authors.affiliation: Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Arizona, AZ, U.S.A.
authors.forename: Aikseng
authors.initials: A
authors.lastname: Ooi
authors.affiliation: Ecole Pratique des Hautes Etudes, Paris and French National Institute of Health and Medical Research U753, Gustave Roussy Cancer Campus, Villejuif, France Faculty of Medicine, Paris-Sud University, Kremlin Bicêtre, Paris, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Richard
authors.affiliation: Department of Biodevices and Diagnostics, Institute of Bioengineering and Nanotechnology, Singapore, Republic of Singapore Division of Medical Oncology, National Cancer Centre Singapore, Singapore, Republic of Singapore minhan.tan@gmail.com teh.bin.tean@singhealth.com.sg.
authors.forename: Min-Han
authors.initials: MH
authors.lastname: Tan
authors.affiliation: Division of Medical Sciences, National Cancer Centre Singapore, Singapore, Republic of Singapore Division of Cancer and Stem Cell Biology, Duke-NUS Graduate Medical School, Singapore, Republic of Singapore minhan.tan@gmail.com teh.bin.tean@singhealth.com.sg.
authors.forename: Bin Tean
authors.initials: BT
authors.lastname: Teh
mesh_terms: D000293:Adolescent; D002292:Carcinoma, Renal Cell / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D005649:Fumarate Hydratase / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D008297:Male
chemical_list: D005649:Fumarate Hydratase
keywords: HLRCC; PRCC2; Warburg effect; bortezomib; fumarate hydratase gene