TBP, PPIA, YWHAZ and EF1A1 are the most stably expressed genes during osteogenic differentiation

Franko, Nina (author), Vrščaj, Lucija Ana (author), Zore, Taja (author), Ostanek, Barbara (author), Marc, Janja (author), Lojk, Jasna (author)

RT-qPCR is the gold standard and the most commonly used method for measuring gene expression. Selection of appropriate reference gene(s) for normalization is a crucial part of RT-qPCR experimental design, which allows accurate quantification and reliability of the results. Because there is no universal reference gene and even commonly used housekeeping genes’ expression can vary under certain conditions, careful selection of an appropriate internal control must be performed for each cell type or tissue and experimental design. The aim of this study was to identify the most stable reference genes during osteogenic differentiation of the human osteosarcoma cell lines MG-63, HOS, and SaOS-2 using the geNorm, NormFinder, and BestKeeper statistical algorithms. Our results show that TBP, PPIA, YWHAZ, and EF1A1 are the most stably expressed genes, while ACTB, and 18S rRNA expressions are most variable. These data provide a basis for future RT-qPCR normalizations when studying gene expression during osteogenic differentiation, for example, in studies of osteoporosis and other bone diseases.

• Language: English
• Keywords: geNorm, reference gene, NormFinder, BestKeeper, osteogenic differentiation, gene expression, osteosarcoma cell line, RT-qPCR
• Work type: Article
• Typology: 1.01 - Original Scientific Article
• Organization: FFA - Faculty of Pharmacy
• Publication status: Published
• Publication version: Version of Record
• Year: 2022
• Number of pages: 17 str.
• Numbering: Vol. 23, iss. 8, art. 4257
• PID: 20.500.12556/RUL-136787
• UDC: 575.117:616-006.34
• ISSN on article: 1422-0067
• DOI: 10.3390/ijms23084257
• COBISS.SI-ID: 105244163
• Publication date in RUL: 20.05.2022

Record is a part of a journal

• Title: International journal of molecular sciences
• Shortened title: Int. j. mol. sci.
• Publisher: MDPI
• ISSN: 1422-0067
• COBISS.SI-ID: 2779162

Licences

• License: CC BY 4.0, Creative Commons Attribution 4.0 International
• Link: http://creativecommons.org/licenses/by/4.0/
• Description: This is the standard Creative Commons license that gives others maximum freedom to do what they want with the work as long as they credit the author.

Secondary language

• Language: Slovenian
• Keywords: referenčni gen, osteogena diferenciacija, celična linija osteosarkoma, gensko izražanje, osteosarkom