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Real-time analysis of gene regulation by glucocorticoid hormones
There is increasing evidence that temporal factors are important in allowing cells to gain additional information from external factors, such as hormones and cytokines. We sought to discover how cell responses to glucocorticoids develop over time, and how the response kinetics vary according to liga...
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BioScientifica
2008
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| 在線閱讀: | https://ncbi.nlm.nih.gov/pmc/articles/PMC2315692/ https://ncbi.nlm.nih.gov/pubmed/18434350 https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1677/JOE-07-0639 |
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pubmed-23156922009-01-27 Real-time analysis of gene regulation by glucocorticoid hormones McMaster, A Chambers, T Meng, Q-J Grundy, S Loudon, A S I Donn, R Ray, D W J Endocrinol Regular papers There is increasing evidence that temporal factors are important in allowing cells to gain additional information from external factors, such as hormones and cytokines. We sought to discover how cell responses to glucocorticoids develop over time, and how the response kinetics vary according to ligand structure and concentration, and hence have developed a continuous gene transcription measurement system, based on an interleukin-6 (IL-6) luciferase reporter gene. We measured the time to maximal response, maximal response and integrated response, and have compared these results with a conventional, end point glucocorticoid bioassay. We studied natural glucocorticoids (corticosterone and cortisol), synthetic glucocorticoids (dexamethasone) and glucocorticoid precursors with weak, or absent bioactivity. We found a close correlation between half maximal effective concentration (EC50) for maximal response, and for integrated response, but with consistently higher EC50 for the latter. There was no relation between the concentration of ligand and the time to maximal response. A comparison between conventional end point assays and real-time measurement showed similar effects for dexamethasone and hydrocortisone, with a less effective inhibition of IL-6 seen with corticosterone. We profiled the activity of precursor steroids, and found pregnenolone, progesterone, 21-hydroxyprogesterone and 17-hydroxyprogesterone all to be ineffective in the real-time assay, but in contrast, progesterone and 21-hydroxyprogesterone showed an IL-6 inhibitory activity in the end point assay. Taken together, our data show how ligand concentration can alter the amplitude of glucocorticoid response, and also that a comparison between real-time and end point assays reveals an unexpected diversity of the function of glucocorticoid precursor steroids, with implications for human disorders associated with their overproduction. BioScientifica 2008-05 /pmc/articles/PMC2315692/ /pubmed/18434350 http://dx.doi.org/10.1677/JOE-07-0639 Text en © 2008 Society for Endocrinology http://www.endocrinology.org/journals/reuselicence/ This is an Open Access article distributed under the terms of the Society for Endocrinology's Re-use Licence (http://www.endocrinology.org/journals/reuselicence/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
| institution |
US National Library of Medicine |
| collection |
PubMed Central |
| language |
English |
| format |
Article |
| topic |
Regular papers |
| spellingShingle |
Regular papers McMaster, A Chambers, T Meng, Q-J Grundy, S Loudon, A S I Donn, R Ray, D W Real-time analysis of gene regulation by glucocorticoid hormones |
| description |
There is increasing evidence that temporal factors are important in allowing cells to gain additional information from external factors, such as hormones and cytokines. We sought to discover how cell responses to glucocorticoids develop over time, and how the response kinetics vary according to ligand structure and concentration, and hence have developed a continuous gene transcription measurement system, based on an interleukin-6 (IL-6) luciferase reporter gene. We measured the time to maximal response, maximal response and integrated response, and have compared these results with a conventional, end point glucocorticoid bioassay. We studied natural glucocorticoids (corticosterone and cortisol), synthetic glucocorticoids (dexamethasone) and glucocorticoid precursors with weak, or absent bioactivity. We found a close correlation between half maximal effective concentration (EC50) for maximal response, and for integrated response, but with consistently higher EC50 for the latter. There was no relation between the concentration of ligand and the time to maximal response. A comparison between conventional end point assays and real-time measurement showed similar effects for dexamethasone and hydrocortisone, with a less effective inhibition of IL-6 seen with corticosterone. We profiled the activity of precursor steroids, and found pregnenolone, progesterone, 21-hydroxyprogesterone and 17-hydroxyprogesterone all to be ineffective in the real-time assay, but in contrast, progesterone and 21-hydroxyprogesterone showed an IL-6 inhibitory activity in the end point assay. Taken together, our data show how ligand concentration can alter the amplitude of glucocorticoid response, and also that a comparison between real-time and end point assays reveals an unexpected diversity of the function of glucocorticoid precursor steroids, with implications for human disorders associated with their overproduction. |
| author |
McMaster, A Chambers, T Meng, Q-J Grundy, S Loudon, A S I Donn, R Ray, D W |
| author_facet |
McMaster, A Chambers, T Meng, Q-J Grundy, S Loudon, A S I Donn, R Ray, D W |
| author_sort |
McMaster, A |
| title |
Real-time analysis of gene regulation by glucocorticoid
hormones |
| title_short |
Real-time analysis of gene regulation by glucocorticoid
hormones |
| title_full |
Real-time analysis of gene regulation by glucocorticoid
hormones |
| title_fullStr |
Real-time analysis of gene regulation by glucocorticoid
hormones |
| title_full_unstemmed |
Real-time analysis of gene regulation by glucocorticoid
hormones |
| title_sort |
real-time analysis of gene regulation by glucocorticoid
hormones |
| publisher |
BioScientifica |
| publisher_facet |
BioScientifica |
| publishDate |
2008 |
| url |
https://ncbi.nlm.nih.gov/pmc/articles/PMC2315692/ https://ncbi.nlm.nih.gov/pubmed/18434350 https://ncbi.nlm.nih.govhttp://dx.doi.org/10.1677/JOE-07-0639 |
| _version_ |
1760537821068656640 |