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EFFECT OF INTERLEUKIN-1β ON GROWTH HORMONE GENE EXPREssION AND ITs POssIBLE MOLECULAR MECHANIsM IN RAT mtt/s sOMATOTROPH cells

引用

Chinese Medical sciences Journal 2008年第4期23卷 193-201页

作者： Feng-ying Gong Jie-ying Deng Yi-fan shi Department of Endocrinology Key Laboratory of Endocrinology of Ministry of Health Peking Union Medical College Hospital Chinese Academy of Medical Sciences ＆ Peking Union Medical College Beijing 100730

Objective To elucidate the effect of interleukin-1β （IL- 1β） on human growth hormone （hGH） gene expression in a rat somatotropic pituitary cell line mtt/s. Methods stably transfected mtt/s cells were firstly established by transfecting 484-Lucl plasmid which contained hGH gene promoter --484 to ＋30 bp and luciferase reporter gene. The effect of IL-1β on hGH gene expression was determined by assaying the luciferase activities. RT-PCR method was also used to determine whether IL-1 recepor mRNA was expressed in mtt/s cells. Results The 10^3 U/mL IL-1β stimulated secretion and synthesis of GH, and promoted the 5＇-promoter activity of GH gene in stably transfected mtt/sGL cells with the action of 1.38 times above the control. Among inhibitors of signaling transduction pathways, mitogen-activated protein kinase kinase （MAPKK/MEK） inhibitor PD98059 （40 μmol/L） and p38 mitogen-activated protein kinase （MAPK） inhibitor sB203580 （5 μmol/L） completely blocked the stimulatory effect of IL-1μ, and phosphatidylinositol-3-kinase （PI3-K） inhibitor LY294002 partly abolished the effect of IL-1μ. Western blot analysis further confirmed the activation of phosphorylated MEK and p38 MAPK in mtt/sGL cells. Neither over-expression of Pit- 1 nor inhibition of Pit- 1 expression affected induction of hGH promoter activity by IL-1μ. A series of deletion constructs of hGH promoter were created to identify the DNA sequence that mediated the effect of IL-1β, and results showed that the stimulatory effect of IL-1β was abolished following deletion of the --196 to -- 132 bp fragment. Conclusions IL-1β promotes GH secretion and synthesis in rat mtt/s somatotroph cells. The stimulatory effect of IL-1β on hGH gene promoter appears to require the activation of MEK, p38 MAPK, PI3-K, and a fragment of promoter sequence that spans the -196 to -132 bp of the gene, but it may be unlinked with Pit-1 protein.

关键词： interleukin-1β growth hormone gene promoter mtt/s cells Pit-1 protein mitogen-activated protein kinase

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MEK AND p38 MAPK-DEPENDANT PATHWAYs ARE INVOLOVED IN THE POsITIVE EFFECT OF INTERLEUKIN-6 ON HUMAN GROWTH HORMONE GENE EXPREssION IN RAT mtt/s sOMATOTROPH cells

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Chinese Medical sciences Journal 2008年第2期23卷 73-80页

Objective To investigate the effect of interleukin-6(IL-6)on the human growth hormone(hGH)gene expression in a rat somatotropic pituitary cell line mtt/*** The plasmids containing various lengths of hGH gene 5'-promoter fragments were *** transfected mtt/s cells were created by cotransfecting the above plasmids and pcDNA3.1(+)with DMRIE-C transfection *** the administration of these cells with IL-6 and/or various inhibitors of signaling transduction pathways,the luciferase activities in mtt/s cells lysis were assayed to demonstrate the effects of IL-6 on hGH gene promoter activity and possibly involved *** The 103 U/mL IL-6 stimulated GH secretion and synthesis,and promoted the 5'-promoter activity of GH gene in stably transfected mtt/sGL cells with the action of 1.69 times above the *** inhibitors of signaling transduction pathways,mitogen-activated protein kinase kinase(MAPKK/MEK)inhibitor PD98059(40 μmol/L)and p38 mitogen-activated protein kinase(MAPK)inhibitor sB203580(5 μmol/L)completely blocked the stimulatory effect of *** blot analysis further confirmed the activation of phosphorylated MEK and p38 MAPK in mtt/sGL *** over-expression of Pit-1 nor inhibition of Pit-1 expression affected IL-6 induction of hGH promoter activity.A series of deletion constructs of hGH promoter were created to identify the DNA sequence that mediated the effect of *** results showed that the stimulatory effect of IL-6 was abolished following deletion of the-196 to-132 bp *** IL-6 promotes GH secretion and synthesis by rat mtt/s somatotroph *** stimulatory effect of IL-6 on hGH gene promoter appears to require the activation of MEK and p38 MAPK,and a fragment of promoter sequence that spans the-196 to-132 bp of the gene,but may be unlinked with Pit-1 protein.

关键词： interleukin-6 growth hormone gene promoter mtt/s cells Pit-1 protein mitogen-activated protein kinase

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