title: Array-based comparative genomic hybridization-guided identification of reference genes for normalization of real-time quantitative polymerase chain reaction assay data for lymphomas, histiocytic sarcomas, and osteosarcomas of dogs.
abstract: OBJECTIVE
  To identify suitable reference genes for normalization of real-time quantitative PCR (RT-qPCR) assay data for common tumors of dogs.
  
  
  METHODS
  Malignant lymph node (n = 8), appendicular osteosarcoma (9), and histiocytic sarcoma (12) samples and control samples of various nonneoplastic canine tissues.
  
  
  METHODS
  Array-based comparative genomic hybridization (aCGH) data were used to guide selection of 9 candidate reference genes. Expression stability of candidate reference genes and 4 commonly used reference genes was determined for tumor samples with RT-qPCR assays and 3 software programs.
  
  
  RESULTS
  LOC611555 was the candidate reference gene with the highest expression stability among the 3 tumor types. Of the commonly used reference genes, expression stability of HPRT was high in histiocytic sarcoma samples, and expression stability of Ubi and RPL32 was high in osteosarcoma samples. Some of the candidate reference genes had higher expression stability than did the commonly used reference genes.
  
  
  CONCLUSIONS
  Data for constitutively expressed genes with high expression stability are required for normalization of RT-qPCR assay results. Without such data, accurate quantification of gene expression in tumor tissue samples is difficult. Results of the present study indicated LOC611555 may be a useful RT-qPCR assay reference gene for multiple tissue types. Some commonly used reference genes may be suitable for normalization of gene expression data for tumors of dogs, such as lymphomas, osteosarcomas, or histiocytic sarcomas.
authors.affiliation: Department of Molecular Biomedical Science, College of Veterinary Medicine, North Carolina State University, Raleigh, NC 27606, USA.
authors.forename: Pei-Chien
authors.initials: PC
authors.lastname: Tsai
authors.forename: Matthew
authors.initials: M
authors.lastname: Breen
mesh_terms: D000818:Animals; D055028:Comparative Genomic Hybridization / Q000379:methods / Q000592:standards / Q000662:veterinary*; D004283:Dog Diseases / Q000235:genetics* / Q000473:pathology; D004285:Dogs; D018628:Gene Dosage; D054747:Histiocytic Sarcoma / Q000235:genetics / Q000473:pathology / Q000662:veterinary*; D007150:Immunohistochemistry; D008223:Lymphoma / Q000235:genetics / Q000473:pathology / Q000662:veterinary*; D012516:Osteosarcoma / Q000235:genetics / Q000473:pathology / Q000662:veterinary*; D012334:RNA, Neoplasm / Q000737:chemistry / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary*; D012015:Reference Standards
chemical_list: D012334:RNA, Neoplasm