PUBLICATION
Development of a Liquid Chromatography/Mass Spectrometry-Based Inhibition Assay for the Screening of Steroid 5-α Reductase in Human and Fish Cell Lines
- Authors
- Kim, D., Cho, H., Eggers, R., Kim, S.K., Ryu, C.S., Kim, Y.J.
- ID
- ZDB-PUB-210511-1
- Date
- 2021
- Source
- Molecules 26(4): (Journal)
- Registered Authors
- Keywords
- 5α-reductase inhibitors, adverse outcome pathway, dihydrotestosterone, dutasteride, finasteride, in vitro
- MeSH Terms
-
- 3-Oxo-5-alpha-Steroid 4-Dehydrogenase/metabolism*
- 5-alpha Reductase Inhibitors/pharmacology*
- Animals
- Calibration
- Cell Line
- Chromatography, Liquid*
- Drug Evaluation, Preclinical/methods*
- Humans
- Mass Spectrometry*
- Oncorhynchus mykiss
- Zebrafish
- PubMed
- 33567691 Full text @ Molecules
Citation
Kim, D., Cho, H., Eggers, R., Kim, S.K., Ryu, C.S., Kim, Y.J. (2021) Development of a Liquid Chromatography/Mass Spectrometry-Based Inhibition Assay for the Screening of Steroid 5-α Reductase in Human and Fish Cell Lines. Molecules. 26(4):.
Abstract
Steroid 5-α reductase (5AR) is responsible for the reduction of steroids to 5-α reduced metabolites, such as the reduction of testosterone to 5-α dihydrotestosterone (DHT). A new adverse outcome pathway (AOP) for 5AR inhibition to reduce female reproduction in fish (AOP 289) is under development to clarify the antiestrogenic effects of 5AR inhibitors in female fish. A sensitive method for the DHT analysis using chemical derivatization and liquid chromatography-tandem mass spectrometry was developed. A cell-based 5AR inhibition assay that utilizes human cell lines, a transient overexpression system, and fish cell lines was developed. The measured IC50 values of two well-known 5AR inhibitors, finasteride and dutasteride, were comparable in the different systems. However, the IC50 of dutasteride in the fish cell lines was lower than that in the human cell lines. Finasteride showed a higher IC50 against the RTG-2 cell line. These results demonstrated that 5ARs inhibition could differ in terms of structural characteristics among species. The assay has high sensitivity and reproducibility and is suitable for the application in 5AR inhibition screening for various endocrine disruption chemicals (EDCs). Future studies will continue to evaluate the quantitative inhibition of 5AR by EDCs to compare the endocrine-disrupting pathway in different species.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping