title: Identification of suitable reference genes for investigating gene expression in human gallbladder carcinoma using reverse transcription quantitative polymerase chain reaction.
abstract: Reverse transcription quantitative polymerase chain reaction (RT‑qPCR) has become a frequently used strategy in gene expression studies. The relative quantification method is an important and commonly used method for the evaluation of RT‑qPCR data. The key aim of this method is to identify an applicable internal reference gene, however, there are currently no suitable reference genes for gene analysis in gallbladder carcinoma. In the present study, screening was performed using 12 common reference genes, which were selected in order to provide an experimental basis for the investigation of gene expression in gallbladder carcinoma. A total of 16 tissue samples of gallbladder carcinoma and their matched normal gallbladder tissues were used. The gene expression stability and applicability of the 12 reference gene candidates were determined using the geNorm, NormFinder and BestKeeper software programs. Following comparison of the results of the three software programs, HPRT1 was identified as the most stably expressed reference gene. In the normal gallbladder group, the relative stably expressed reference gene was PPIA and in the entire sample group, the relatively stably expressed reference gene was PPIA. The present study also demonstrated that the combination of the three reference genes was the most appropriate. The recommended combinations were PPIA + PUM1 + ACTB for the total sample group, GAPDH + PBGD + ALAS1 for the gallbladder carcinoma group and PPIA + PUM1 + TBP for the paired normal gallbladder group.
authors.affiliation: Department of Neurology, China‑Japan Union Hospital, Jilin University, Changchun, Jilin 130033, P.R. China.
authors.forename: Shan
authors.initials: S
authors.lastname: Yu
authors.affiliation: Central Laboratory, Second Hospital, Jilin University, Changchun, Jilin 130041, P.R. China.
authors.forename: Qiwei
authors.initials: Q
authors.lastname: Yang
authors.affiliation: Department of Hepatopancreatobiliary Surgery, China‑Japan Union Hospital, Jilin University, Changchun, Jilin 130033, P.R. China.
authors.forename: Jing Hui
authors.initials: JH
authors.lastname: Yang
authors.affiliation: Central Laboratory, Second Hospital, Jilin University, Changchun, Jilin 130041, P.R. China.
authors.forename: Zhenwu
authors.initials: Z
authors.lastname: Du
authors.affiliation: Central Laboratory, Second Hospital, Jilin University, Changchun, Jilin 130041, P.R. China.
authors.forename: Guizhen
authors.initials: G
authors.lastname: Zhang
mesh_terms: D000328:Adult; D000368:Aged; D002277:Carcinoma / Q000235:genetics* / Q000473:pathology; D005260:Female; D005706:Gallbladder Neoplasms / Q000235:genetics* / Q000473:pathology; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D008875:Middle Aged; D060787:Neoplasm Grading; D009367:Neoplasm Staging; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics; D020871:RNA Stability; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D016271:Proto-Oncogene Proteins c-myc