Transient agonist-induced regulation of the cholecystokinin-A and cholecystokinin-B receptor mRNA levels in rat pancreatic acinar AR42J cells

Günther, Rainer; Carstens, Ove; Schmidt, Wolfgang E.; Fölsch, Ulrich R.

doi:10.1159/000069142

Cited by 6 publications

(7 citation statements)

References 31 publications

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“…The Cck2r was highly expressed in the presence of CHX which indicates that Cck2r is a primary gastrin-induced gene and negatively feedback-regulated at the mRNA level (data not shown). This is in accordance with findings by others showing that gastrin increases CCK2R expression at the mRNA and protein levels, both in cell cultures [54,55] and in vivo [55]. More investigation is needed to address the status of the gastrin receptor upon different duration of gastrin treatment.…”

Section: Resultssupporting

confidence: 87%

“…Furthermore, AR42J cells express gastrin receptors endogenously and can therefore be used as a model system to study gastrin responses like proliferation [65], differentiation [66] and apoptosis [24-27]. In the time-series experiment we treated cells with 10 nM gastrin in accordance with several other studies investigating gastrin-induced responses in the AR42J cell line [25,54,67]. The cells were grown in 6-well plates (3 ×10 5 cells/well) for 72 h. Then the growth medium was replenished with 2 ml serum-free DMEM, and the cells serum starved for 20–24 h before adding gastrin.…”

Section: Methodsmentioning

confidence: 99%

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The duration of gastrin treatment affects global gene expression and molecular responses involved in ER stress and anti-apoptosis

et al. 2013

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BackgroundHow cells decipher the duration of an external signal into different transcriptional outcomes is poorly understood. The hormone gastrin can promote a variety of cellular responses including proliferation, differentiation, migration and anti-apoptosis. While gastrin in normal concentrations has important physiological functions in the gastrointestine, prolonged high levels of gastrin (hypergastrinemia) is related to pathophysiological processes.ResultsWe have used genome-wide microarray time series analysis and molecular studies to identify genes that are affected by the duration of gastrin treatment in adenocarcinoma cells. Among 403 genes differentially regulated in transiently (gastrin removed after 1 h) versus sustained (gastrin present for 14 h) treated cells, 259 genes upregulated by sustained gastrin treatment compared to untreated controls were expressed at lower levels in the transient mode. The difference was subtle for early genes like Junb and c-Fos, but substantial for delayed and late genes. Inhibition of protein synthesis by cycloheximide was used to distinguish between primary and secondary gastrin regulated genes. The majority of gastrin upregulated genes lower expressed in transiently treated cells were primary genes induced independently of de novo protein synthesis. This indicates that the duration effect of gastrin treatment is mainly mediated via post-translational signalling events, while a smaller fraction of the differentially expressed genes are regulated downstream of primary transcriptional events. Indeed, sustained gastrin treatment specifically induced prolonged ERK1/2 activation and elevated levels of the AP-1 subunit protein JUNB. Enrichment analyses of the differentially expressed genes suggested that endoplasmic reticulum (ER) stress and survival is affected by the duration of gastrin treatment. Sustained treatment exerted an anti-apoptotic effect on serum starvation-induced apoptosis via a PKC-dependent mechanism. In accordance with this, only sustained treatment induced anti-apoptotic genes like Clu, Selm and Mcl1, while the pro-apoptotic gene Casp2 was more highly expressed in transiently treated cells. Knockdown studies showed that JUNB is involved in sustained gastrin induced expression of the UPR/ER stress related genes Atf4, Herpud1 and Chac1.ConclusionThe duration of gastrin treatment affects both intracellular signalling mechanisms and gene expression, and ERK1/2 and AP-1 seem to play a role in converting different durations of gastrin treatment into distinct cellular responses.

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Section: Resultssupporting

confidence: 87%

Section: Methodsmentioning

confidence: 99%

The duration of gastrin treatment affects global gene expression and molecular responses involved in ER stress and anti-apoptosis

et al. 2013

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“…Gastrin has been reported to upregulate CCK2R expression in the gastric surface mucus cell line GSM06 ( Nakajima et al 2002 ) and in pancreatic AR42J cells ( Gunther et al 2003 ), in support of much earlier reports that CCK2R expression in vivo might be regulated by serum gastrin concentrations ( Takeuchi et al 1979 , 1980 ). We used CCK2R promoter–reporter constructs to determine whether gastrin could induce CCK2R transcription in AGS-G R cells and in the untransformed cell line RGM1.…”

Section: Resultssupporting

confidence: 75%

Regulation of mammalian gastrin/CCK receptor (CCK2R) expression in vitro and in vivo

Ashurst

Varró

Dimaline

2007

Experimental Physiology

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The gastrin/CCK receptor (CCK2R) mediates the physiological functions of gastrin in the stomach, including stimulation of acid secretion and cellular proliferation and migration, but little is known about the factors that regulate its expression. We identified endogenous CCK2R expression in several cell lines and used luciferase promoter-reporter constructs to define the minimal promoter required for transcription in human gastric adenocarcinoma, AGS, and rat gastric mucosa, RGM1, cells. Consensus binding sites for SP1, C/EBP and GATA were essential for activity. Following serum withdrawal from RGM1 and AR42J cells, endogenous CCK2R mRNA abundance and the activity of a CCK2R promoter-reporter construct were significantly elevated. Transcription of CCK2R was also increased in AGS-G R and RGM1 cells by gastrin through mechanisms partly dependent upon protein kinase C (PKC) and mitogen/extracellular signal-regulated kinase (MEK). Gastrin significantly increased endogenous CCK2R expression in RGM1 cells, and CCK2R protein expression was elevated in the stomach of hypergastrinaemic animals. In mice with cryoulcers in the acid-secreting mucosa, CCK2R expression increased progressively in the regenerating mucosa adjacent to the ulcer repair margin, evident at 6 days postinjury and maximal at 13 days. De novo expression of CCK2R was observed in the submucosa beneath the repairing ulcer crater 6-9 days postinjury. Many of the cells in mucosa and submucosa that expressed CCK2R in response to cryoinjury were identified as myofibroblasts, since they coexpressed vimentin and smooth muscle α-actin but not desmin. The data suggest that increased CCK2R expression might influence the outcome of epithelial inflammation or injury and that the response may be mediated in part by myofibroblasts.

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“…The results presented here support the latter explanation. This kind of homologous regulation of receptor expression has been shown for several GPCRs (Siegrist et al 1994;Schanstra et al 1998;Froidevaux and Eberle 2002;Gunther et al 2003). Both NPFF and NPFF2 receptor co-exist within the same sites in the CNS (Yang and Iadarola 2003;Zeng et al 2003) and hence, the homologous regulation of the receptor is also possible in vivo.…”

Section: Discussionmentioning

confidence: 64%

Regulation of endogenous human NPFF2 receptor by neuropeptide FF in SK‐N‐MC neuroblastoma cell line

Änkö

Panula

2005

Journal of Neurochemistry

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Neuropeptide FF has many functions both in the CNS and periphery. Two G protein-coupled receptors (NPFF1 and NPFF2 receptors) have been identified for neuropeptide FF. The expression analysis of the peptide and receptors, together with pharmacological and physiological data, imply that NPFF2 receptor would be the primary receptor for neuropeptide FF. Here, we report for the first time a cell line endogenously expressing hNPFF2 receptor. These SK-N-MC neuroblastoma cells also express neuropeptide FF. We used the cells to investigate the hNPFF2 receptor function. The pertussis toxin-sensitive inhibition of adenylate cyclase activity upon receptor activation indicated coupling to G i/o proteins. Upon agonist exposure, the receptors were internalized and the mitogen-activated protein kinase cascade was activated.Upon neuropeptide FF treatment, the actin cytoskeleton was reorganized in the cells. The expression of hNPFF2 receptor mRNA was up-regulated by neuropeptide FF. Concomitant with the receptor mRNA, the receptor protein expression was increased. The homologous regulation of hNPFF2 receptor correlates with our previous results in vivo showing that during inflammation, the up-regulation of neuropeptide FF mRNA precedes that of NPFF2 receptor. The regulation of hNPFF2 receptor by NPFF could also be important in the periphery where neuropeptide FF has been suggested to function as a hormone.

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Transient agonist-induced regulation of the cholecystokinin-A and cholecystokinin-B receptor mRNA levels in rat pancreatic acinar AR42J cells

Cited by 6 publications

References 31 publications

The duration of gastrin treatment affects global gene expression and molecular responses involved in ER stress and anti-apoptosis

The duration of gastrin treatment affects global gene expression and molecular responses involved in ER stress and anti-apoptosis

Regulation of mammalian gastrin/CCK receptor (CCK2R) expression in vitro and in vivo

Regulation of endogenous human NPFF2 receptor by neuropeptide FF in SK‐N‐MC neuroblastoma cell line

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