title: Modulation of the circadian clock by glucocorticoid receptor isoforms in the H295R cell line.
abstract: Peripheral clocks are set by different nervous, hormonal and metabolic stimuli, and regulate the circadian expression of several genes. We investigated whether a peripheral clock could be induced in the human adrenocortical cell line H295R and whether glucocorticoid receptor isoforms (GRα and GRß) are involved in this clock system. After synchronization of cells with serum shock, the rhythmic oscillation of clock genes PER1, PER2, REV-ERBα, and ARNTL was confirmed. In addition, H295R cells even without serum shock showed rhythmic expression of PER1, PER2, CRY1 and ARNTL. Glucocorticoid treatment induced a rapid response of PER1, PER2 and CRY1 in a GRα-dependent manner. Continuous glucocorticoid stimulation after 6h caused suppression of REV-ERBα. Administration of a GR antagonist, RU486, disrupted the circadian oscillation of clock genes and prevented the acute changes in PER1, PER2 and CRY1 levels. Overexpression of the GRß isoform alone did not alter the expression of the examined clock genes, but did prevent the GRα-related suppression of REV-ERBα. These alterations occurred independently from ACTH and CRH. Our data demonstrate that a peripheral clock system is present in a human adrenocortical cell line and that periodic oscillations of clock genes are influenced by glucocorticoids, mainly through GRα.
authors.affiliation: 2nd Department of Medicine, Faculty of Medicine, Semmelweis University, Budapest, Hungary; Hungarian Academy of Sciences-Semmelweis University "Lendulet" Hereditary Endocrine Tumors Research Group, Budapest, Hungary.
authors.forename: Zsolt
authors.initials: Z
authors.lastname: Nagy
authors.affiliation: 2nd Department of Medicine, Faculty of Medicine, Semmelweis University, Budapest, Hungary.
authors.forename: Alexa
authors.initials: A
authors.lastname: Marta
authors.affiliation: Hungarian Academy of Sciences-Semmelweis University Molecular Medicine Research Group, Budapest, Hungary.
authors.forename: Henriett
authors.initials: H
authors.lastname: Butz
authors.affiliation: Hungarian Academy of Sciences-Semmelweis University "Lendulet" Hereditary Endocrine Tumors Research Group, Budapest, Hungary.
authors.forename: Istvan
authors.initials: I
authors.lastname: Liko
authors.affiliation: 2nd Department of Medicine, Faculty of Medicine, Semmelweis University, Budapest, Hungary; Hungarian Academy of Sciences-Semmelweis University Molecular Medicine Research Group, Budapest, Hungary.
authors.forename: Karoly
authors.initials: K
authors.lastname: Racz
authors.affiliation: Hungarian Academy of Sciences-Semmelweis University "Lendulet" Hereditary Endocrine Tumors Research Group, Budapest, Hungary; Department of Laboratory Medicine Institute, Faculty of Medicine, Semmelweis University, Budapest, Hungary. Electronic address: patocs.attila@med.semmelweis-univ.hu.
authors.forename: Attila
authors.initials: A
authors.lastname: Patocs
mesh_terms: D056930:ARNTL Transcription Factors / Q000235:genetics / Q000378:metabolism; D002460:Cell Line; D057906:Circadian Clocks / Q000187:drug effects / Q000235:genetics; D002940:Circadian Rhythm / Q000187:drug effects / Q000235:genetics; D056931:Cryptochromes / Q000235:genetics / Q000378:metabolism; D005938:Glucocorticoids / Q000494:pharmacology; D006801:Humans; D056950:Period Circadian Proteins / Q000235:genetics / Q000378:metabolism; D011965:Receptors, Glucocorticoid / Q000235:genetics / Q000378:metabolism*
chemical_list: D056930:ARNTL Transcription Factors; C538963:CRY1 protein, human; D056931:Cryptochromes; D005938:Glucocorticoids; C539022:PER1 protein, human; C421902:PER2 protein, human; D056950:Period Circadian Proteins; D011965:Receptors, Glucocorticoid; C411470:glucocorticoid receptor alpha; C121943:glucocorticoid receptor beta
keywords: Adrenal gland; Circadian rhythm; Clock genes; Glucocorticoid receptor; H295R cell line; Peripheral clock