Abstract
BackgroundRetinoid Receptors are involved in development and cell homeostasis. Alterations of their expressions have been observed in lung cancer. However, retinoid chemoprevention trials in populations at risk to develop such tumors have failed. Therefore, the pertinence of new clinical trials using second generation retinoid requires prior better understanding of retinoid signalling. This is our aim when validating extensively research tools, focused on Retinoic Acid Receptor beta, whose major role in lung cancer is documented.MethodsBiocomputing was used to assess the genomic organization of RAR beta. Its putative RAR-beta1' promoter features were investigated experimentally. Specific measures realized, with qRT-PCR Syber Green assays and a triplex of Taqman probes, were extensively validated to establish Retinoid Receptors mRNAs reference values for in vivo normal human bronchial cells, lung tumors and cell lines. Finally, a pan-RAR-beta antibody was generated and extensively validated by western-blot and immunoprecipitation.ResultsNo promoter-like activity was found for RAR-beta1'. RAR-beta2 mRNAs increase signs the normal differentiation of the human bronchial epithelium while a decrease is observed in most lung cancer cell lines. Accordingly, it is also, along with RXR beta, down-regulated in lung tumors. When using nuclear extracts of BEAS-2B and normal lung cells, only the RAR-beta2 long protein isoform was recognized by our antibody.ConclusionRigorous samples processing and extensive biocomputing, were the key factors for this study. mRNA reference values and validated tools can now be used to advance researches on retinoid signalling in the lung.
Highlights
Retinoid Receptors are involved in development and cell homeostasis
We used qRT-PCR analysis to measure RRs mRNAs expressions in several samples and compared the results obtained in Syber Green assays to those given by a triplex of Taqman probes that was conceived to save invaluable mRNA samples
Such transcripts were not detected in the samples tested and no activity was found for RAR-beta1'
Summary
Retinoid Receptors are involved in development and cell homeostasis Alterations of their expressions have been observed in lung cancer. The pertinence of new clinical trials using second generation retinoid requires prior better understanding of retinoid signalling This is our aim when validating extensively research tools, focused on Retinoic Acid Receptor beta, whose major role in lung cancer is documented. Vitamin A and its active derivatives referred to as retinoids are non-steroid hormones which play a critical role in the development and homeostasis of vertebrate tissues [1,2,3] They exert their actions by regulating the expression of target genes, influencing likewise: cell proliferation, differentiation and apoptosis through nuclear Retinoid Receptors (RRs) binding [4]. This underlines the need for more knowledge about retinoid signalling in the normal human lung
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
