Change search
ReferencesLink to record
Permanent link

Direct link
Endogenous Gene and Protein Expression of Drug-Transporting Proteins in Cell Lines Routinely Used in Drug Discovery Programs
KTH, School of Biotechnology (BIO), Proteomics.ORCID iD: 0000-0001-6990-1905
Show others and affiliations
2009 (English)In: Drug Metabolism And Disposition, ISSN 0090-9556, E-ISSN 1521-009X, Vol. 37, no 12, 2275-2283 p.Article in journal (Refereed) Published
Abstract [en]

The aim of this study was to investigate the gene and protein expression profiles of important drug-transporting proteins in human cell lines commonly used for studies of drug transport mechanisms. Human cell lines used to transiently or stably express single transporters [HeLa, human embryonic kidney (HEK) 293] and leukemia cell lines used to study drug resistance by ATP-binding cassette transporters (HL-60, K562) were investigated and compared with organotypic cell lines (HepG2, Saos-2, Caco-2, and Caco-2 TC7). For gene expression studies, real-time polymerase chain reaction was used, whereas monospecific polyclonal antibodies were generated and used to investigate protein expression by immunohistochemistry. Thirty-six transporters were studied for gene expression, and nine were studied for protein expression. The antibodies were validated using expression patterns in human tissues. Finally, the function of one ubiquitously expressed transporter, MCT1/SLC16A1, was investigated using [C-14]lactic acid as a substrate. In general, the adherent cell lines (HeLa, HEK293) displayed low transporter expression, and the expression patterns were barely affected by transfection. The leukemia cell lines (K562, HL-60) and Saos-2 also had low endogenous transporter expression, whereas the organotypic cell lines (HepG2 and Caco-2) showed higher expression of some transporters. Comparison of gene and protein expression profiles gave poor correlations, but better agreement was obtained for antibodies with a good validation score, indicating that antibody quality was a significant variable. It is noteworthy that the monocarboxylic acid-transporting protein MCT1 was significantly expressed in all and was functional in most of the cell lines, indicating that MCT1 may be a confounding factor when the transport of small anionic drugs is investigated.

Place, publisher, year, edition, pages
2009. Vol. 37, no 12, 2275-2283 p.
Keyword [en]
human tumor-cells, multidrug-resistance, caco-2 cells, genomic, organization, efflux transporters, human intestine, p-glycoprotein, human tissues, in-vitro, mrp2
URN: urn:nbn:se:kth:diva-18970DOI: 10.1124/dmd.109.028654ISI: 000271935200002ScopusID: 2-s2.0-71049160053OAI: diva2:337017
QC 20100525Available from: 2010-08-05 Created: 2010-08-05 Last updated: 2011-02-14Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textScopus

Search in DiVA

By author/editor
Al-Khalili Szigyarto, CristinaUhlén, Mathias
By organisation
In the same journal
Drug Metabolism And Disposition

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 53 hits
ReferencesLink to record
Permanent link

Direct link