title: Validation of internal reference genes for relative quantitation studies of gene expression in human laryngeal cancer.
abstract: BACKGROUND
  The aim of this study was to determine the expression stabilities of 12 common internal reference genes for the relative quantitation analysis of target gene expression performed by reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) in human laryngeal cancer.
  
  
  METHODS
  Hep-2 cells and 14 laryngeal cancer tissue samples were investigated. The expression characteristics of 12 internal reference gene candidates (18S rRNA, GAPDH, ACTB, HPRT1, RPL29, HMBS, PPIA, ALAS1, TBP, PUM1, GUSB, and B2M) were assessed by RT-qPCR. The data were analyzed by three commonly used software programs: geNorm, NormFinder, and BestKeeper.
  
  
  RESULTS
  The use of the combination of four internal reference genes was more appropriate than the use of a single internal reference gene. The optimal combination was PPIA + GUSB + RPL29 + HPRT1 for both the cell line and tissues; while the most appropriate combination was GUSB + RPL29 + HPRT1 + HMBS for the tissues.
  
  
  CONCLUSIONS
  Our recommended internal reference genes may improve the accuracy of relative quantitation analysis of target gene expression performed by the RT-qPCR method in further gene expression research on laryngeal tumors.
authors.affiliation: Department of Stomatology, China-Japan Union Hospital, Jilin University , Changchun , China.
authors.forename: Xiaofeng
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Neurology, China-Japan Union Hospital, Jilin University , Changchun , China.
authors.forename: Jinting
authors.initials: J
authors.lastname: He
authors.affiliation: Department of Stomatology, China-Japan Union Hospital, Jilin University , Changchun , China.
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, Second Hospital, Jilin University , Changchun , China.
authors.forename: Ming
authors.initials: M
authors.lastname: Ren
authors.affiliation: Department of Anesthesiology, China-Japan Union Hospital, Jilin University , Changchun , China.
authors.forename: Sujie
authors.initials: S
authors.lastname: Gao
authors.affiliation: Department of Nephrology, China-Japan Union Hospital, Jilin University , Changchun , China.
authors.forename: Guanjie
authors.initials: G
authors.lastname: Zhao
authors.affiliation: Department of Orthopedics, Second Hospital, Jilin University , Changchun , China.
authors.forename: Jincheng
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Orthopedics, Second Hospital, Jilin University, Changchun, China; Central Laboratory of Second Hospital, Jilin University, Changchun, China.
authors.forename: Qiwei
authors.initials: Q
authors.lastname: Yang
keywords: Expression stability; Housekeeping gene; Human laryngeal cancer; Normalization; Reverse transcription quantitative polymerase chain reaction