title: Non-malignant respiratory epithelial cells preferentially proliferate from resected non-small cell lung cancer specimens cultured under conditionally reprogrammed conditions.
abstract: The "conditionally reprogrammed cells" (CRC) method, using a Rho kinase inhibitor and irradiated mouse fibroblast cells has been described for the efficient growth of cells from malignant and non-malignant samples from primary tumor and non-malignant sites. Using the CRC method, four institutions independently cultured tumor tissues from 48 non-small cell lung cancers (NSCLC, mostly from primary resected tumors) and 22 non-malignant lungs. We found that epithelial cells could be cultured from tumor and non-malignant lung. However, epithelial cells cultured from tumors had features of non-malignant respiratory epithelial cells which include: 1) among 22 mutations found in the original tumors only two mutations were found in the CRC cultures with reduced frequency (31% to 13% and 92% to 15% from original tumor and CRC culture respectively); 2) copy number variation was analyzed in 9 tumor and their CRC cultures and only diploid patterns were found in CRC cultures; 3) mRNA expression profiles were similar to those of normal respiratory epithelial cells; and 4) co-culture of tumor and non-malignant lung epithelial cells resulted in mostly non-malignant cells. We conclude that CRC method is a highly selective and useful method for the growth of non-malignant respiratory epithelial cells from tumor specimens and only occasionally do such CRC cultures contain a small subpopulation of cancer cells marked by oncogenic mutations. While our findings are restricted to resected primary NSCLC, they indicated the necessity to fully characterize all CRC cultures and the need to develop culture technology that facilitates the growth of primary lung cancers.
authors.affiliation: Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|The Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Boning
authors.initials: B
authors.lastname: Gao
authors.affiliation: Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|The Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Chunxian
authors.initials: C
authors.lastname: Huang
authors.affiliation: Division of Thoracic Surgery, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Kemp
authors.initials: K
authors.lastname: Kernstine
authors.affiliation: Department of Pathology, Yale University, New Haven, CT, USA.
authors.forename: Vasiliki
authors.initials: V
authors.lastname: Pelekanou
authors.affiliation: Department of Pathology, Yale University, New Haven, CT, USA.
authors.forename: Yuval
authors.initials: Y
authors.lastname: Kluger
authors.affiliation: Interdepartmental Program in Computational Biology and Bioinformatics, Yale University, New Haven, CT, USA.
authors.forename: Tingting
authors.initials: T
authors.lastname: Jiang
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Jennifer R
authors.initials: JR
authors.lastname: Peters-Hall
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Melissa
authors.initials: M
authors.lastname: Coquelin
authors.affiliation: Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|The Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Luc
authors.initials: L
authors.lastname: Girard
authors.affiliation: Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|The Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|The Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Kenneth
authors.initials: K
authors.lastname: Huffman
authors.affiliation: Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Dwight
authors.initials: D
authors.lastname: Oliver
authors.affiliation: Department of Thoracic Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, USA.
authors.forename: Fumi
authors.initials: F
authors.lastname: Kinose
authors.affiliation: Department of Thoracic Oncology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, USA.
authors.forename: Eric
authors.initials: E
authors.lastname: Haura
authors.affiliation: Department of Biostatistics and Bioinformatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, USA.
authors.forename: Jamie K
authors.initials: JK
authors.lastname: Teer
authors.affiliation: Department of Drug Discovery, H. Lee Moffitt Cancer Center and Research Institute, Tampa, Florida, USA.
authors.forename: Uwe
authors.initials: U
authors.lastname: Rix
authors.affiliation: Department of Medicine, Division of Medical Oncology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA.
authors.forename: Anh T
authors.initials: AT
authors.lastname: Le
authors.affiliation: Department of Pathology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA.
authors.forename: Dara L
authors.initials: DL
authors.lastname: Aisner
authors.affiliation: Department of Medicine, Division of Medical Oncology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA.|Department of Pathology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA.
authors.forename: Marileila
authors.initials: M
authors.lastname: Varella-Garcia
authors.affiliation: Department of Medicine, Division of Medical Oncology, University of Colorado Anschutz Medical Campus, Aurora, Colorado, USA.
authors.forename: Robert C
authors.initials: RC
authors.lastname: Doebele
authors.affiliation: Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Kyle R
authors.initials: KR
authors.lastname: Covington
authors.affiliation: Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Oliver A
authors.initials: OA
authors.lastname: Hampton
authors.affiliation: Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Harsha V
authors.initials: HV
authors.lastname: Doddapaneni
authors.affiliation: Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Joy C
authors.initials: JC
authors.lastname: Jayaseelan
authors.affiliation: Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Jianhong
authors.initials: J
authors.lastname: Hu
authors.affiliation: Human Genome Sequencing Center, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: David A
authors.initials: DA
authors.lastname: Wheeler
authors.affiliation: Department of Cell Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Jerry W
authors.initials: JW
authors.lastname: Shay
authors.affiliation: Department of Pathology, Yale University, New Haven, CT, USA.
authors.forename: David L
authors.initials: DL
authors.lastname: Rimm
authors.affiliation: Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|The Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Adi
authors.initials: A
authors.lastname: Gazdar
authors.affiliation: Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|The Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Pharmacology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Internal Medicine, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: John D
authors.initials: JD
authors.lastname: Minna
mesh_terms: D000072283:A549 Cells; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000473:pathology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000235:genetics*; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods*; D056915:DNA Copy Number Variations; D004252:DNA Mutational Analysis / Q000379:methods; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism*; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods; D020022:Genetic Predisposition to Disease / Q000235:genetics; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009154:Mutation; D020545:Respiratory Mucosa / Q000166:cytology / Q000378:metabolism; D014407:Tumor Cells, Cultured
keywords: cell culture; conditionally reprogrammed cells; non-small cell lung cancer; respiratory epithelial cells; rock inhibitor