The effect of 16,16-dimethyl prostaglandin E2 on proliferation of an intestinal goblet cell line and its synthesis and secretion of mucin glycoproteins

doi:10.1016/0952-3278(93)90047-Z

Prostaglandins, Leukotrienes and Essential Fatty Acids

Volume 48, Issue 6, June 1993, Pages 423-428

https://doi.org/10.1016/0952-3278(93)90047-Z Get rights and content

Abstract

The effect of 16,16′-dimethyl prostaglandin E² (dmPGE₂) on the human colonic adenocarcinoma derived mucus-secreting goblet cell line HT29-18N2 was investigated. The proliferation rate of HT29-18N2 was increased by exposure to 10 or 100 μM dmPGE₂. Exposure to 10 or 100 μM dmPGE₂ caused a significant decrease in the rate of radiolabeled glucosamine incorporation into newly synthesized glycoproteins during an 8 or 24 h exposure. At concentrations as low as 1 μM, dmPGE₂ accelerated the secretion of mucin glycoproteins as assessed by the release of newly synthesized radiolabeled glycoproteins, a mucin-specific enzyme-linked immunoassay and a whole-mount immunofluorescence assay. A 1 h exposure to dmPGE₂ did not, however, result in a morphometrically detectable decrease in intracellular mucous granule stores or elicit any other readily detectable morphological change. The experimental results suggest elevated levels of PGs may contribute to the previously recognized decreases in intracellular mucin stores and shifts in the types of mucins species present at sites of mucosal inflammation in ulcerative colitis patients.

References (29)

A Robert et al.
Cytoprotection by prostaglandins in rats
Gastroenterology
(1979)
ER Lacy et al.
Microscopic analysis of ethanol damage to rat gastric mucosa after treatment with a prostaglandin
Gastroenterology
(1982)
JL Wallace et al.
Role of mucus in the repair of gastric epithelial damage in the rat
Gastroenterology
(1986)
AJ Uribe et al.
Initial kinetic changes of prostaglandin E2-induced hyperplasia of the rat small intestinal epithelium occur in the villous compartments
Gastroenterology
(1988)
TE Phillips et al.
Human intestinal goblet cells inmonolayer culture: characterization of a mucus-secreting subclone derived from the HT29 colon adenocarcinoma cell line
Gastroenterology
(1988)
GNJ Tytgat et al.
Influence of 15(R)-15-methyl prostaglandin E2 on human gastric mucosa. A light microscopic, cell kinetic, and ultrastructural study
Gastroenterology
(1986)
U Seidler et al.
Effects of endogenous and exogenous prostaglandins on glycoprotein synthesis and secretion in isolated rabbit gastric mucosa
Gastroenterology
(1988)
T Wahlin et al.
Effects of intraluminal prostaglandin E2 in vivo on secretory behavior and ultrastructural changes in mouse gallbladder epithelium
Gastroenterology
(1988)
DK Podolsky et al.
Glycoprotein composition of colonic mucosa: specific alterations in ulcerative colitis
Gastroenterology
(1984)
JL Wallace et al.
Prostaglandin protection of rat colonic mucosa from damage by ethanol
Dig Dis Sci
(1985)

JL Wallace et al.

Reduction by cytoprotective agents of ethanol-induced damage to the rat gastric mucosa: a correlated morphological and physiological study

Can J Physiol Pharmacol

(1982)

A Allen et al.

Mucus and mucosal protection

GP Morris et al.

Mechanisms of mucosal recovery from acute gastric damage: roles of extracellular mucus and cell migration

RA Goodlad et al.

Effects of the prostaglandin analogue misoprostol on cell proliferation in the canine small intestine

Exp Physiol

(1991)

Cited by (28)

KAG-308, a newly-identified EP<inf>4</inf>-selective agonist shows efficacy for treating ulcerative colitis and can bring about lower risk of colorectal carcinogenesis by oral administration
2015, European Journal of Pharmacology
Citation Excerpt :
EP4 is strongly expressed in the goblet cells of the rat gastrointestinal tract, and its expression is increased in colonic mucosa during colitis development (Nitta et al., 2002; Northey et al., 2000). PGE2 is suggested to induce the proliferation of goblet cells (Phillips et al., 1993). Signaling via EP4 is also shown to stimulate colonic mucin exocytosis in human colonic epithelial cells and rat colon (Belley and Chadee, 1999).
Agonists for EP₄ receptor, a prostaglandin E₂ receptor subtype, appear to be a promising therapeutic strategy for ulcerative colitis (UC) due to their anti-inflammatory and epithelial regeneration activities. However, the clinical development of orally-available EP₄ agonists for mild to moderate UC has not yet been reported. Furthermore, the possibility of an increased risk of colitis-associated cancer (CAC) through direct proliferative effects on epithelial cells via EP₄ signaling has not been ruled out. Recently, we identified KAG-308 as an orally-available EP₄-selective agonist. Here, we investigated the pharmacological and pharmacokinetic profiles of KAG-308. Then, we compared KAG-308 and sulfasalazine (SASP) for their abilities to prevent colitis and promote mucosal healing in a mouse model of dextran sulfate sodium (DSS)-induced colitis. Finally, the effect of KAG-308 treatment on CAC was evaluated in an azoxymethane (AOM)/DSS-induced CAC mouse model. KAG-308 selectively activated EP₄ and potently inhibited tumor necrosis factor-α production in peripheral whole blood and T cells. Oral administration of KAG-308, which showed relatively high bioavailability, suppressed the onset of DSS-induced colitis and promoted histological mucosal healing, while SASP did not. KAG-308 also prevented colorectal carcinogenesis by inhibiting colitis development and consequently decreasing mortality in a CAC model, whereas SASP had marginal effects. In contrast, MF-482, an EP₄ antagonist, increased mortality. These results indicated that orally-administered KAG-308 suppressed colitis development and promoted mucosal healing. Moreover, it exhibited preventive effects on colorectal carcinogenesis, and thus may be a new therapeutic strategy for the management of UC that confers a reduced risk of colorectal carcinogenesis.
Chemotherapy-induced mucositis: The role of mucin secretion and regulation, and the enteric nervous system
2013, NeuroToxicology
Citation Excerpt :
A 1 hour exposure of 16,16′-dimethyl prostaglandin E2 (dmPGE2) on the human colonic adenocarcinoma derived mucus-secreting goblet cell line (HT29-18N2) was shown to be associated with an increase in the proliferation rate of HT29-18N2 cells, and accelerated the secretion of mucin glycoproteins, as determined by the release of newly synthesized radiolabeled glycoproteins (Phillips et al., 1993). However, no morphological changes were observed (Phillips et al., 1993). Studies in a rat model of mucositis have shown an increase in cavitated goblet cells and mucin secretion in response to chemotherapy, suggesting PGE2 may play a role in the increased mucin secretion associated with the development of mucositis (Stringer et al., 2009a,b).
Alimentary mucositis is a severe, dose-limiting, toxic side effect of cytotoxic chemotherapy and radiotherapy. Patients with mucositis often have reductions or breaks imposed on cytotoxic therapy, which may lead to reduced survival. Furthermore, there is an increased risk of infection and hospitalization, compounding the cost of treatment. There are currently limited therapeutic options for mucositis, and no effective prevention available. Mucin expression and secretion have been shown to be associated with mucositis. Furthermore, mucins exhibit protective effects on the alimentary tract through reducing mechanical and chemical stress, preventing bacterial overgrowth and penetration, and digestion of the mucosa. Additionally, a number of studies have implicated some key neurotransmitters in both mucositis and mucin secretion, suggesting that the enteric nervous system may also play a key role in the development of mucositis.
The cyclo-oxygenase-2 inhibitor, rofecoxib, attenuates mucosal damage due to colitis induced by trinitrobenzene sulphonic acid in rats
2003, European Journal of Pharmacology
Cyclo-oxygenase-2 overexpression has been described in experimental colitis. However, there are controversial findings suggesting that its inhibition by selective cyclo-oxygenase-2 inhibitors not only may have a beneficial effect on experimental colitis, but also exacerbate the inflammation-associated colonic injury. Thus, the role of cyclo-oxygenase-2 inhibitors in the possible modulation of colon inflammation is controversial and remains uncertain. In this study, we evaluated the effects of the selective cyclo-oxygenase-2 inhibitor, rofecoxib, on the extent and severity of ulcerative colitis caused by intracolonic administration of 2,4,6-trinitrobenzene sulphonic acid (TNBS) in rats. The lesions and the inflammatory response were assessed by histology and measurement of myeloperoxidase activity. Interleukin-1β, prostaglandin E₂ and D₂ levels in colon mucosa and the immunohistochemical expression of the cyclo-oxygenases-1 and -2 were also studied. Finally, we investigated the effects of rofecoxib on apoptosis of colonocytes by the appearance of DNA fragmentation. Inflammation following TNBS was characterized by increased colonic wall thickness, oedema, diffuse inflammatory cell infiltration in the mucosa, and necrosis. Increased myeloperoxidase activity, as an index of neutrophil infiltration in the mucosa, and interleukin-1β levels were also measured in the colon. Administration of rofecoxib significantly (P<0.05) reduced the colonic damage, the degree of neutrophil infiltration, and interleukin-1β levels. In addition, apoptosis was significantly increased in TNBS-treated rats, but not in rofecoxib plus TNBS-treated rats. We concluded that rofecoxib seems to have beneficial effects in TNBS-induced colitis by diminishing the initial stage of inflammation, probably by a mechanism related to inhibition of prostaglandin E₂ by the cyclo-oxygenase-2 pathway. The data suggest that cyclo-oxygenase-2-selective inhibitors may have a therapeutic role in ulcerative colitis.
Molecular cloning and characterization of a novel UDP-GlcNAc: GalNAc-peptide β1,3-N-acetylglucosaminyltransferase (β3Gn-T6), an enzyme synthesizing the core 3 structure of O-glycans
2002, Journal of Biological Chemistry
The core 3 structure of the O-glycan, GlcNAcβ1–3GalNAcα1-serine/threonine, an important precursor in the biosynthesis of mucin-type glycoproteins, is synthesized by UDP-N-acetylglucosamine:GalNAc-peptide β1,3-N- acetylglucosaminyltransferase (β3Gn-T; core 3 synthase). The core 3 structure is restricted in its occurrence to mucins from specific tissues such as the stomach, small intestine, and colon. A partial sequence encoding a novel member of the human β3Gn-T family was found in one of the data bases. We cloned a complementary DNA of this gene and named it β3Gn-T6. The putative amino acid sequence of β3Gn-T6 retains the β3Gn-T motifs and is predicted to comprise a typical type II membrane protein. The soluble form of β3Gn-T6 expressed in insect cells showed β3Gn-T activity toward GalNAcα-p-nitrophenyl and GalNAcα1-serine/threonine. The β1,3-linkage between GlcNAc and GalNAc of the enzyme reaction product was confirmed by high performance liquid chromatography and NMR analyses. β3Gn-T6 effectively transferred a GlcNAc to the GalNAc residue on MUC1 mucin, resulting in the synthesis of a core 3 structure. Real time PCR analysis revealed that the β3Gn-T6 transcript was restricted in its distribution, mainly to the stomach, colon, and small intestine. We concluded that β3Gn-T6 is the most logical candidate for the core 3 synthase, which plays an important role in the synthesis of mucin-type O-glycans in digestive organs.
Cellular distribution of prostanoid EP receptors mRNA in the rat gastrointestinal tract
2000, Prostaglandins and Other Lipid Mediators
The inhibition of PGE₂ synthesis resulting from sustained NSAIDs therapy has been linked to gastrointestinal irritations and ulceration. The multiple physiological effects of PGE₂ in the gut are mediated through the activation of four receptors termed EP_1–4. The aim of the study was to determine the precise distribution of the four prostaglandin E₂ receptors in the rat stomach, small intestine, and colon. We used non-radioactive in situ hybridization techniques on paraffin-embedded tissue. Mucous cells of the stomach and goblet cells of the small intestine and colon were found to express mRNA for all four EP subtypes. A positive hybridization signal for EP₁, EP₃, and EP₄ was detected in the parietal cells of the stomach whereas the chief cells expressed low levels of EP₁ and EP₃. The EP₁ and EP₃ receptor mRNA could also be detected in the muscularis mucosa, longitudinal muscle and enteric ganglias of the stomach and small intestine. However, close examination of the enteric ganglias indicated that most of the positive labeling was localized to the glial cells, although some neurons did express EP₃. In conclusion, we have detailed the distribution of prostanoid EP receptors in the gut at the cellular level, giving new insights to the role of prostaglandins in gastrointestinal functions.
A novel biological role for prostaglandin D<inf>2</inf> is suggested by distribution studies of the rat DP prostanoid receptor
1999, European Journal of Pharmacology
We report the cloning, functional expression and cell-specific localization of the rat homologue of the prostaglandin D₂ receptor (DP). In situ hybridization, utilizing multiple digoxigenin-labelled riboprobes and their complementary sense controls, was performed to determine the detailed distribution of DP receptor mRNA in the central nervous system and the gastrointestinal tract. Within the brain, the leptomeninges and choroid plexus expressed DP receptor mRNA. Transcripts detected in the spinal cord were localized to the sensory and motor neurons of the dorsal and ventral horns, respectively, suggesting a role for the DP receptor in the modulation of central nervous system processes, including pain transmission. Within the gastrointestinal tract (stomach, duodenum, ileum and colon) signals were highly localized to the mucous-secreting goblet cells and the columnar epithelium. These findings suggest a novel biological role for prostaglandin D₂-mediated activity at the DP receptor, namely mucous secretion. In addition, radioligand binding assays (saturation analyses and equilibrium competition assays) and functional assays (measuring cAMP accumulation) were performed to characterize the recombinant rat DP receptor expressed in human embryonic kidney (HEK) 293(EBNA) cells. A single site of binding (K_D=14 nM, B_max=115 fmol/mg protein) was measured for prostaglandin D₂-specific binding to the rat DP receptor. Prostaglandin D₂ proved to be a potent agonist at the rat DP receptor (EC₅₀=5 nM). The rank order of efficacy for DP receptor specific agonists [prostaglandin D₂=prostaglandin J₂=BW 245C (5-(6-carboxyhexyl)-1-(3-cyclohexyl-3-hydroxypropylhydantoin))>L-644,698 ((4-(3-(3-(3-hydroxyoctyl)-4-oxo-2-thiazolidinyl) propyl) benzoic acid) (racemate)] reflected the affinity with which the ligands bound to the receptor.

View all citing articles on Scopus

View full text

The effect of 16,16-dimethyl prostaglandin E2 on proliferation of an intestinal goblet cell line and its synthesis and secretion of mucin glycoproteins

Abstract

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Gastroenterology

Prostaglandin protection of rat colonic mucosa from damage by ethanol

Dig Dis Sci

Reduction by cytoprotective agents of ethanol-induced damage to the rat gastric mucosa: a correlated morphological and physiological study

Can J Physiol Pharmacol

Mucus and mucosal protection

Mechanisms of mucosal recovery from acute gastric damage: roles of extracellular mucus and cell migration

Effects of the prostaglandin analogue misoprostol on cell proliferation in the canine small intestine

Exp Physiol

The effect of 16,16-dimethyl prostaglandin E₂ on proliferation of an intestinal goblet cell line and its synthesis and secretion of mucin glycoproteins