Validation of internal reference genes for relative quantitation studies of gene expression in human laryngeal cancer

PeerJ

article

Xiaofeng Wang¹  Jinting He²  Wei Wang¹  Ming Ren³  Sujie Gao⁴  Guanjie Zhao⁵  Jincheng Wang³  Qiwei Yang³ 

[1] Department of Stomatology, China–Japan Union Hospital, Jilin University;Department of Neurology, China–Japan Union Hospital, Jilin University;Department of Orthopedics, Second Hospital, Jilin University;Department of Anesthesiology, China–Japan Union Hospital, Jilin University;Department of Nephrology, China–Japan Union Hospital, Jilin University;Central Laboratory of Second Hospital, Jilin University

关键词: Housekeeping gene;    Reverse transcription quantitative polymerase chain reaction;    Human laryngeal cancer;    Normalization;    Expression stability;   

DOI  :  10.7717/peerj.2763

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

BackgroundThe 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.MethodsHep-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.ResultsThe 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.ConclusionsOur 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.

【 授权许可】

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