Cyclooxygenase 1 mRNA expression is undetectable in Madin Darby Canine Kidney cells
- DOI
- Language of the publication
- English
- Date
- 2015-03-24
- Type
- Article
- Author(s)
- Pelletier, Guillaume
- Padhi, Bhaja K.
- Publisher
- BMC
Abstract
Background: Madin Darby Canine Kidney (MDCK) cells form polarized epithelium in vitro and are routinely used in research fields ranging from protein trafficking to influenza. However, the canine origin of these cells also means that compared to man or mouse, genomic resources are more limited and performance of commercially available antibodies often untested. The synthesis of pro-inflammatory prostaglandins in the kidney is mediated by the constitutively expressed cyclooxygenase 1 and the inducible cyclooxygenase 2 (COX-1 and COX-2, respectively). There are conflicting reports on the expression of COX-1 and COX-2 in MDCK cells and this lingering uncertainty about such important pharmacological targets may affect the interpretation of results obtained from this cell line. Results: In order to definitively settle the issue of cyclooxygenase expression in MDCK cells, we designed PCR primers based on dog genomic sequences to probe COX-1 and COX-2 mRNA expression in MDCK cells and dog kidney. We report that while COX-1 and COX-2 genes are both expressed in dog kidney, COX-1 expression is undetectable in MDCK cells. Conclusions: By improving the characterization of cyclooxygenase expression in MDCK cells, this study will contribute to a better understanding of the properties of this cell line and lead to improved experimental designs and data interpretations.
Plain language summary
Established cell lines are useful tools for in vitro toxicity studies. However, proper interpretation of the results obtained from these cells requires a good understanding of their underlying biology. Expression of the continually active and inducible forms of a gene involved in inflammation (namely, COX-1 and COX-2) is generally assumed in Madin Darby Canine Kidney (MDCK) cells, despite conflicting reports. This lingering uncertainty can affect the interpretation of a wide range of assays where MDCK cells are used. In order to better characterize the biological properties of a cell line that may hold promise in toxicity studies, Health Canada scientists took advantage of the sequencing of the dog genome to assess the expression of these two genes by the use of the molecular biology technique polymerase chain reaction (PCR). It was found that while both COX-1 and COX-2 gene expression can be detected in the dog kidney, the continually active gene COX-1 is not in fact expressed in MDCK cells. A better characterization and understanding of the MDCK cell line will lead to improved design of studies using this model, to better interpretation of the results obtained, and ultimately to a better understanding of which in vitro models have the most promise for deployment in toxicity studies aimed at characterizing human health risks.
Subject
- Health,
- Health and safety