Journal
JOURNAL OF ENZYME INHIBITION AND MEDICINAL CHEMISTRY
Volume 36, Issue 1, Pages 581-592Publisher
TAYLOR & FRANCIS LTD
DOI: 10.1080/14756366.2021.1882451
Keywords
cancer cells; indoleamine 2; 3-dioxygenase activity; kynurenine; tryptophan metabolites; glycation
Funding
- European Union from European Regional Development Fund under the Operational Programme Development of Eastern Poland 2007-2013 [POPW.01.03.00-06-003/09-00]
- Polish National Science Centre, OPUS13 grant [2017/25/B/NZ4/01198]
Ask authors/readers for more resources
A new HPLC-DAD method for determining IDO1 activity in human cancer cells has been developed and successfully validated, demonstrating its applicability in different cancer cell types. This approach provides a useful model for studying the role of the kynurenine pathway in cancer biology.
The kynurenine pathway is the major tryptophan degradation routes generating bioactive compounds important in physiology and diseases. Depending on cell type it is initiated enzymatically by tryptophan-2,3-dioxygenase (TDO) or indoleamine-2,3-dioxygenase 1 and 2 (IDO1 and IDO2) to yield N-formylkynurenine as the precursor of further metabolites. Herein, we describe an accurate high-pressure liquid chromatography coupled with a diode array detector (HPLC-DAD) method to serve for IDO1 activity determination in human cancer cells cultured in vitro. Enzymatic activity was expressed as the rate of x29f;-kynurenine generation by 1 mg of proteins obtained from cancer cells. Our approach shows the limit of detection and limit of quantification at 12.9 and 43.0 nM Kyn, respectively. Applicability of this method was demonstrated in different cells (ovarian and breast cancer)exposed to various conditions and has successfully passed the validation process. This approach presents a useful model to study the role of kynurenine pathway in cancer biology.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available