Back to Search
Start Over
Validation of Suitable Housekeeping Genes for the Normalization of mRNA Expression for Studying Tumor Acidosis
- Source :
- International Journal of Molecular Sciences, Vol 19, Iss 10, p 2930 (2018), International Journal of Molecular Sciences, Volume 19, Issue 10
- Publication Year :
- 2018
- Publisher :
- MDPI AG, 2018.
-
Abstract
- Similar to other types of cancer, acidification of tumor microenvironment is an important feature of osteosarcoma, and a major source of cellular stress that triggers cancer aggressiveness, drug resistance, and progression. Among the different effects of low extracellular pH on tumor cells, we have recently found that short-term exposure to acidosis strongly affects gene expression. This alteration might also occur for the most commonly used housekeeping genes (HKG), thereby causing erroneous interpretation of RT-qPCR data. On this basis, by using osteosarcoma cells cultured at different pH values, we aimed to identify the ideal HKG to be considered in studies on tumor-associated acidosis. We verified the stability of 15 commonly used HKG through five algorithms (NormFinder, geNorm, BestKeeper, &Delta<br />CT, coefficient of variation) and found that no universal HKG is suitable, since at least four HKG are necessary for proper normalization. Furthermore, according to the acceptable range of values, YWHAZ, GAPDH, GUSB, and 18S rRNA were the most stable reference genes at different pH. Our results will be helpful for future investigations focusing on the effect of altered microenvironment on cancer behavior, particularly on the effectiveness of anticancer therapies in acid conditions.
- Subjects :
- 0301 basic medicine
Reproducibility of Result
acidosi
Validation Studies as Topic
Biology
Article
Catalysis
lcsh:Chemistry
Inorganic Chemistry
03 medical and health sciences
Neoplasms
Reference genes
Gene expression
medicine
Humans
tumor microenvironment
RNA, Messenger
Physical and Theoretical Chemistry
lcsh:QH301-705.5
Molecular Biology
Spectroscopy
housekeeping gene
Tumor microenvironment
Genes, Essential
Gene Expression Profiling
RT-qPCR
Organic Chemistry
Reproducibility of Results
Computational Biology
Cancer
General Medicine
Reference Standards
Hydrogen-Ion Concentration
medicine.disease
Computer Science Applications
Housekeeping gene
Gene Expression Regulation, Neoplastic
Gene expression profiling
030104 developmental biology
lcsh:Biology (General)
lcsh:QD1-999
YWHAZ
Cancer research
Neoplasm
Reference Standard
Osteosarcoma
acidosis
Human
Subjects
Details
- ISSN :
- 14220067
- Volume :
- 19
- Database :
- OpenAIRE
- Journal :
- International Journal of Molecular Sciences
- Accession number :
- edsair.doi.dedup.....e85468a56d5cc9e4ce390745020f5dd4
- Full Text :
- https://doi.org/10.3390/ijms19102930