title: Interleukin-6 increases the expression of key proteins associated with steroidogenesis in human NCI-H295R adrenocortical cells.
abstract: Mechanisms of interleukin-6 (IL-6)-induced cortisol release (CR) were investigated by exposing H295R cells to IL-6 and determining mRNA/protein expression (PCR/western blots) for steroidogenic enzymes (SE), steroidogenic acute regulatory protein (StAR), steroidogenic factor-1 (SF-1) (enhances SE/StAR expression), activator protein 1 (AP-1) (regulates SE/StAR expression) and adrenal hypoplasia congenita-like protein (DAX-1) (inhibits SE/StAR expression). Promoter activity of StAR (SPA) was measured by a luciferase-coupled promoter. Cortisol release was increased by 10ng/mL IL-6 (24h P<0.01). Proteins/mRNAs (StAR, cholesterol side chain cleavage enzyme, SF-1, AP-1) and SPA were increased by IL-6 (60min 1-50ng/mL IL-6; 5ng/mL IL-6 30-120min P<0.05). Four other SE proteins/mRNAs were also increased by 10ng/mL IL-6 (60min P<0.01). Protein/mRNA for DAX-1 was decreased by IL-6 (60min 1-50ng/mL IL-6; 5ng/mL IL-6 30-120min P<0.01). Phosphorylation of Janus kinase (JAK) and signal transducer and activator of transcription (STAT) was increased by IL-6 (JAK2 60min 1-50ng/mL IL-6; 10ng/mL IL-6 5-60min P<0.05; STAT1 and STAT3 60min 10ng/mL IL-6 P<0.01). Inhibition of JAK/STAT with AG490 (10μM) or piceatannol (50μM) blocked (P<0.01 10ng/mL IL-6vs. IL-6 plus AG490 or piceatannol) IL-6-induced increases in SPA and StAR mRNA. In summary, IL-6-induced CR may be facilitated by increased StAR and SE mediated by increased SF-1 and AP-1, decreased DAX-1, and increased phosphorylation of JAK/STAT.
authors.affiliation: Department of Physiology and Developmental Biology and Neuroscience Center, 2025 LSB, Brigham Young University, Provo, UT 84602, United States.
authors.forename: Janae
authors.initials: J
authors.lastname: Strickland
authors.affiliation: Department of Physiology and Developmental Biology and Neuroscience Center, 2025 LSB, Brigham Young University, Provo, UT 84602, United States.
authors.forename: Stephen
authors.initials: S
authors.lastname: McIlmoil
authors.affiliation: Department of Physiology and Developmental Biology and Neuroscience Center, 2025 LSB, Brigham Young University, Provo, UT 84602, United States.
authors.forename: Brice J
authors.initials: BJ
authors.lastname: Williams
authors.affiliation: Department of Physiology and Developmental Biology and Neuroscience Center, 2025 LSB, Brigham Young University, Provo, UT 84602, United States.
authors.forename: Dennis C
authors.initials: DC
authors.lastname: Seager
authors.affiliation: Department of Physiology and Developmental Biology and Neuroscience Center, 2025 LSB, Brigham Young University, Provo, UT 84602, United States.
authors.forename: James P
authors.initials: JP
authors.lastname: Porter
authors.affiliation: Department of Physiology and Developmental Biology and Neuroscience Center, 2025 LSB, Brigham Young University, Provo, UT 84602, United States. Electronic address: Allan_Judd@BYU.EDU.
authors.forename: Allan M
authors.initials: AM
authors.lastname: Judd
mesh_terms: D000302:Adrenal Cortex / Q000187:drug effects / Q000378:metabolism; D015153:Blotting, Western; D002460:Cell Line; D006801:Humans; D006854:Hydrocortisone / Q000378:metabolism; D015850:Interleukin-6 / Q000494:pharmacology*; D053612:Janus Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D012333:RNA, Messenger / Q000378:metabolism; D050791:STAT Transcription Factors / Q000378:metabolism*; D054339:Steroidogenic Factor 1 / Q000378:metabolism; D013256:Steroids / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism
chemical_list: D015850:Interleukin-6; D012333:RNA, Messenger; D050791:STAT Transcription Factors; D054339:Steroidogenic Factor 1; D013256:Steroids; D014157:Transcription Factors; D053612:Janus Kinases; D006854:Hydrocortisone
keywords: DAX-1; Interleukin-6; JAK/STAT pathway; NCI-H295R adrenocortical cells; Steroidogenesis; Steroidogenic factor-1