Effect of Myosin Light Chain Kinase Inhibitor Wortmannin on the Wound Repair of Cultured Gastric Mucosal Cells

Watanabe, Sumio; Wang, Xian‐En; Hirose, Miyoko; Sato, Nobuhiro

doi:10.1006/bbrc.1994.1300

Cited by 27 publications

(13 citation statements)

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“…Wound-healing experiments were done as described by Watanabe et al (28) with modifications. Following TGF-h1 or BMP7 treatment, medium was removed and a standardized, artificial wound was made by mechanical denudation with a rotating tip fitted with a 10 AL pipette propelled by a small electric drill (MB186; Minicraft, Greenville, WI).…”

Section: Functional Assaysmentioning

confidence: 99%

In vivo and In vitro Evidence for Transforming Growth Factor-β1-Mediated Epithelial to Mesenchymal Transition in Esophageal Adenocarcinoma

Rees

Onwuegbusi²,

Save³

et al. 2006

Cancer Research

104

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There is increasing evidence that epithelial to mesenchymal transition (EMT) is involved in cancer progression. Because local invasion and metastasis occurs early in the pathogenesis of esophageal adenocarcinoma, we hypothesized that EMT may be important in this disease. Using immunohistochemistry in a well-characterized set of adenocarcinoma tissues, we showed down-regulation of epithelial markers (E-cadherin and cytokeratin 18) and up-regulation of mesenchymal markers (vimentin and A-smooth muscle actin) with concomitant transforming growth factor-B1 (TGF-B1) expression at the invasive margin compared with the central tumor. A panel of esophageal cell lines was examined for the ability of TGF-B1 to induce EMT in vitro. TE7 cells were selected as a model because TGF-B1 (0-5 ng/mL) treatment induced morphologic and molecular expression changes suggestive of EMT. In TE7 cells, these TGF-B1-induced changes were reversed by 100 ng/ mL of bone morphogenetic protein 7 (BMP7), another member of the TGF-B1 superfamily. EMT was mediated via canonical TGF-B1 signaling with concomitant up-regulation of SMAD-interacting protein 1. Alterations in functional variables (aggregation, wounding, motility, and invasion) following TGF-B1 treatment were consistent with a more invasive phenotype. These functional changes were reversed by BMP7 and SMAD4 RNA interference in vitro. These data suggest that TGF-B1-mediated EMT may be relevant in esophageal carcinogenesis. (Cancer Res 2006; 66(19): 9583-90)

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Section: Functional Assaysmentioning

confidence: 99%

In vivo and In vitro Evidence for Transforming Growth Factor-β1-Mediated Epithelial to Mesenchymal Transition in Esophageal Adenocarcinoma

Rees

Onwuegbusi²,

Save³

et al. 2006

Cancer Research

104

View full text Add to dashboard Cite

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“…[10][11][12][13][14][15][16] As we reported previously, primary cultured gastric epithelial cells used in this model consisted mainly of mucous cells. 10 In this model, after an artificial wound was made, epithelial cells located on the wound edge formed lamellipodia at the leading edge of the individual cells.…”

Section: Discussionmentioning

confidence: 99%

“…As we reported previously, the majority (more than 90%) of cells in the monolayer cell sheet were periodic acid-Schiff (PAS) staining-positive mucous cells. [10][11][12] Following the formation of a complete monolayer cell sheet, a circular artificial wound with a cell-free area of constant size (2 mm 2 ) was created in the center of the dish with a pencil-type mixer with a rotating silicon tip (diameter, 1 mm) without damaging the dish surface, as described previously. 10 All of the following restoration assays, including the cell migration assay and the proliferation assay, were performed under serum-free conditions.…”

Section: Total Gastric Epithelial Cell Restorationmentioning

confidence: 99%

“…[10][11][12] Briefly, in the first group, BrdU (10 µg/ml) was added to the medium immediately after wounding and the cells incubated for 12 h, and then cells were processed for immunostaining. In the second group, BrdU was added 12 h after wounding, and the incubation was continued for an additional 12 h. In the third group, BrdU was added 24 h after wounding; and in the fourth group, it was added 36 h after wounding.…”

Section: Proliferation Assaymentioning

confidence: 99%

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Effects of teprenone on gastric epithelial restoration in a rabbit cultured cell model

Watanabe

Wang

Hirose

et al. 1998

Journal of Gastroenterology

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The mechanism of action of gastrocytoprotective agents is not fully understood. We assessed the effects of an anti-ulcer agent, teprenone, and bile acid on epithelial restoration. Teprenone with or without deoxycholic acid was added to a complete confluent cultured gastric epithelial cell sheet after wounding. Restoration was monitored for 48 h, and the wound size was assessed. Migration velocity was measured, and proliferation was detected by sequential staining with bromodeoxyuridine. The labeling index was calculated. Restoration was completed within 48h in controls, whereas deoxycholic acid retarded repair. The migration velocity was suppressed by deoxycholic acid treatment. The number of proliferative cells peaked at 36 h (labeling index, 1.7%) in controls. In the deoxycholic acid group, the maximal labeling index was 0.5% at 48 h. Teprenone abolished the bile acid-induced retardation. Teprenone showed cytoprotective effects against deoxycholic acid-induced inhibition of epithelial cell migration and proliferation.

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“…Japanese white rabbits (2.0 kg, male) were used. Rabbit gastric epithelial cells were isolated according to methods described previously 5 , 6 . Briefly, stomachs were removed from anaesthetized (Nembutal; 50 mg/kg, i.p.)…”

Section: Methodsmentioning

confidence: 99%

The novel histamine H₂ receptor antagonist FRG‐8813 prevents delay of wound repair induced by hydrogen peroxide in a rabbit gastric epithelial cell system

Sato

Watanabe

Wang

et al. 1998

J of Gastro and Hepatol

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The effects of a novel histamine H2 receptor antagonist (FRG‐8813) on the restoration process of gastric epithelial wounds were assessed using an in vitro wound healing model. FRG‐8813 (1, 10 mol/L) was added to a complete confluent monolayer cell sheet after artificial wounding. The restoration process was analysed by a time‐lapse video system and cell migration, proliferation and apoptosis were assessed. Hydrogen peroxide (1, 3 mmol/L) inhibited restoration after wounding by suppressing cell migration and proliferation and induced epithelial cell apoptosis around the wound. The addition of FRG‐8813 abolished the hydrogen peroxide‐induced retardation and prevented apoptosis, although FRG‐8813 itself did not enhance wound healing. FRG‐8813 may act as a radical scavenger as well as having an anti‐secretory action and may have favourable effects on peptic ulcer healing.

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Effect of Myosin Light Chain Kinase Inhibitor Wortmannin on the Wound Repair of Cultured Gastric Mucosal Cells

Cited by 27 publications

References 0 publications

In vivo and In vitro Evidence for Transforming Growth Factor-β1-Mediated Epithelial to Mesenchymal Transition in Esophageal Adenocarcinoma

In vivo and In vitro Evidence for Transforming Growth Factor-β1-Mediated Epithelial to Mesenchymal Transition in Esophageal Adenocarcinoma

Effects of teprenone on gastric epithelial restoration in a rabbit cultured cell model

The novel histamine H₂ receptor antagonist FRG‐8813 prevents delay of wound repair induced by hydrogen peroxide in a rabbit gastric epithelial cell system

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Effect of Myosin Light Chain Kinase Inhibitor Wortmannin on the Wound Repair of Cultured Gastric Mucosal Cells

Cited by 27 publications

References 0 publications

In vivo and In vitro Evidence for Transforming Growth Factor-β1-Mediated Epithelial to Mesenchymal Transition in Esophageal Adenocarcinoma

In vivo and In vitro Evidence for Transforming Growth Factor-β1-Mediated Epithelial to Mesenchymal Transition in Esophageal Adenocarcinoma

Effects of teprenone on gastric epithelial restoration in a rabbit cultured cell model

The novel histamine H2 receptor antagonist FRG‐8813 prevents delay of wound repair induced by hydrogen peroxide in a rabbit gastric epithelial cell system

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The novel histamine H₂ receptor antagonist FRG‐8813 prevents delay of wound repair induced by hydrogen peroxide in a rabbit gastric epithelial cell system