Design, optimization and validation of genes commonly used in expression studies on DMH/AOM rat colon carcinogenesis model

View/ Open
Issue date
2019-01-29Author
Riera-Escamilla, Antoni
Gou, Gemma
Suggested citation
Bars-Cortina, David;
Riera-Escamilla, Antoni;
Gou, Gemma;
Piñol Felis, Carme;
Motilva Casado, Mª José;
.
(2019)
.
Design, optimization and validation of genes commonly used in expression studies on DMH/AOM rat colon carcinogenesis model.
PeerJ, 2019, vol. 7, e6372, p. 1-18.
https://doi.org/10.7717/peerj.6372.
Metadata
Show full item recordAbstract
Colorectal cancer (CRC), also known as colon cancer, is the third most common form of cancer worldwide in men and the second in women and is characterized by several genetic alterations, among them the expression of several genes. 1,2-dimethylhydrazine (DMH) and its metabolite azoxymethane (AOM) are procarcinogens commonly used to induce colon cancer in rats (DMH/AOM rat model). This rat model has been used to study changes in mRNA expression in genes involved in this pathological condition. However, a lack of proper detailed PCR primer design in the literature limits the reproducibility of the published data. The present study aims to design, optimize and validate the qPCR, in accordance with the MIQE (Minimum Information for Publication of Quantitative Real-Time PCR Experiments) guidelines, for seventeen genes commonly used in the DMH/AOM rat model of CRC (Apc, Aurka, Bax, Bcl2, β-catenin, Ccnd1, Cdkn1a, Cox2, Gsk3beta, IL-33, iNOs, Nrf2, p53, RelA, Smad4, Tnfα and Vegfa) and two reference genes (Actb or β-actin and B2m). The specificity of all primer pairs was empirically validated on agarose gel, and furthermore, the melting curve inspection was checked as was their efficiency (%) ranging from 90 to 110 with a correlation coefficient of r2 > 0.980. Finally, a pilot study was performed to compare the robustness of two candidate reference genes.
Is part of
PeerJ, 2019, vol. 7, e6372, p. 1-18European research projects
Collections
Except where otherwise noted, this item's license is described as cc-by (c) Bars-Cortina et al., 2019
Related items
Showing items related by title, author, creator and subject.
-
Application of in vitro gastrointestinal digestion and colonic fermentation models to pomegranate products (juice, pulp and peel extract) to study the stability and catabolism of phenolic compounds
Mosele, Juana; Macià i Puig, Ma Alba; Romero Fabregat, Mª Paz; Motilva Casado, Mª José; Rubió Piqué, Laura (John Wiley & Sons, 2015)Pomegranate fruit contains a wide range of phenolic compounds that have been related to several health benefits. The stability of pomegranate phenols during digestion was tested by the application of in vitro gastrointestinal ... -
Optimization and validation of analytical methods for the simultaneous determination of antioxidants: Application to the analysis of tomato sauces.
Motilva Casado, Mª José; Macià i Puig, Ma Alba; Romero Fabregat, Mª Paz; Labrador, Agustín; Domínguez, Alba; Peiró, Lluís (Elsevier, 2014)In the present study, simultaneous extraction of natural antioxidants (phenols and carotenoids) in complex matrices, such as tomato sauces, is presented. The tomato sauce antioxidant compounds studied were the phenolics ... -
Study of the catabolism of thyme phenols combining in-vitro fermentation and human intervention.
Mosele, Juana; Martín Peláez, Sandra María; Macià i Puig, Ma Alba; Farràs, Marta; Valls, Rosa M.; Catalán Santos, Úrsula; Motilva Casado, Mª José (American Chemical Society, 2014)The gut metabolism of four thyme phenolics (monoterpenes thymol and carvacrol, rosmarinic acid, and eriodictyol) was evaluated in vitro. After the in vitro transformations of the individual phenols had been studied, the ...