Induction of Highly Proliferative Phenotypes in Cultured Glomerular Mesangial Cells by Benzo[a]pyrene Alone or in Combination with Methoxamine

Bowes, Russell C.; Weber, Thomas J.; Ramos, Kenneth S.

doi:10.1006/abbi.1995.9968

Cited by 13 publications

(8 citation statements)

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“…Electrophoretic Mobility Shift Assays-Electrophoretic mobility shift assays were carried out after the method of Denison et al (33) as modified by Bowes et al (34). Briefly, cells were rinsed twice and lysed in ice-cold HEGD (25 mM HEPES, pH 7.6, 1.5 mM EDTA, 10% glycerol, 1 mM dithiothreitol, 0.1 mg/ml phenylmethylsulfonyl fluoride, 0.75 mM spermidine, 0.15 mM spermine) by homogenization.…”

Section: Methodsmentioning

confidence: 99%

Acrolein Causes Inhibitor κB-independent Decreases in Nuclear Factor κB Activation in Human Lung Adenocarcinoma (A549) Cells

Horton

Biswal

Corrigan

et al. 1999

Journal of Biological Chemistry

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Acrolein is a highly electrophilic ␣,␤-unsaturated aldehyde to which humans are exposed in various situations. In the present study, the effects of sublethal doses of acrolein on nuclear factor B (NF-B) activation in A549 human lung adenocarcinoma cells were investigated. Immediately following a 30-min exposure to 45 fmol of acrolein/cell, glutathione (GSH) and DNA synthesis and NF-B binding were reduced by more than 80%. All parameters returned to normal or supranormal levels by 8 h post-treatment. Pretreatment with acrolein completely blocked 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced activation of NF-B. Cells treated for 1 h with 1 mM diethyl maleate (DEM) showed a 34 and 53% decrease in GSH and DNA synthesis, respectively. DEM also reduced NF-B activation by 64% at 2 h posttreatment, with recovery to within 22% of control at 8 h. Both acrolein and DEM decreased NF-B function ϳ50% at 2 h after treatment with TPA, as shown by a secreted alkaline phosphatase reporter assay. GSH returned to control levels by 8 h after DEM treatment, but proliferation remained significantly depressed for 24 h. Interestingly, DEM caused a profound decrease in NF-B binding, even at doses as low as 0.125 mM that had little effect on GSH. Neither acrolein nor DEM had any effect on the levels of phosphorylated or nonphosphorylated inhibitor B-␣ (IB-␣). Furthermore, acrolein decreased NF-B activation in cells depleted of IB-␣ by TPA stimulation in the presence of cycloheximide, demonstrating that the decrease in NF-B activation was not the result of increased binding by the inhibitory protein. This conclusion was further supported by the finding that acrolein modified NF-B in the cytosol prior to chemical dissociation from IB with detergent. Together, these data support the conclusion that the inhibition of NF-B activation by acrolein and DEM is IB-independent. The mechanism appears to be related to direct modification of thiol groups in the NF-B subunits.

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Section: Methodsmentioning

confidence: 99%

Acrolein Causes Inhibitor κB-independent Decreases in Nuclear Factor κB Activation in Human Lung Adenocarcinoma (A549) Cells

Horton

Biswal

Corrigan

et al. 1999

Journal of Biological Chemistry

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“…[ 3 H]thymidine incorporation into DNA Growth factor-induced changes in proliferation were also evaluated by [ 3 H]thymidine incorporation into DNA (Bowes et al, 1995). Cells were incubated with 0.5 Ci [ 3 H]thymidine/ml for 24 h. Collagen was digested as described above.…”

Section: Assays Of Relative Apoptotic Events In Rptesmentioning

confidence: 99%

Hepatocyte growth factor induces tubulogenesis of primary renal proximal tubular epithelial cells

et al. 1999

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Hepatocyte growth factor (HGF)-induced tubulogenesis has been demonstrated with renal epithelial cell lines grown in collagen gels but not with primary cultured renal proximal tubular epithelial cells (RPTEs). We show that HGF selectively induces proliferation and branching morphogenesis of primary cultured rat RPTEs. Additional growth factors including fibroblast growth factor (FGF)-1, epidermal growth factor (EGF), FGF-7, or insulin-like growth factor-1 (IGF-1) did not selectively induce tubulogenesis. However, when administered in combination, these factors initiated branching morphogenesis comparable to HGF alone and greatly augmented HGF-induced proliferation and branching. Microscopic analysis revealed that branching RPTEs were undergoing tubulogenesis and formed a polarized epithelium. TGF-beta1 blocked HGF- or growth factor cocktail (GFC; HGF, FGF-1, EGF, IGF-1)-induced proliferation and branching morphogenesis. Adding TGF-beta1 after GFC-induced tubulogenesis had occurred caused a progressive regression of the tubular structures, a response associated with an increase in apoptosis of the RPTEs. Primary cultured RPTEs are capable of undergoing HGF-induced tubulogenesis. Unlike cell lines, combinations of growth factors differentially augment the response.

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“…Electrophoretic mobility shift assay (EMSA). EMSAs were carried out as described previously (6). LLC-PK 1 cells were collected and lysed in buffer A [25 mM HEPES, pH 7.6, 1.5 mM EDTA, 10% glycerol, 1 mM dithiothreitol (DTT), and 0.1 mg/ml phenylmethylsulfonyl fluoride] using 20 strokes with a Dounce homogenizer.…”

Section: Chemicalsmentioning

confidence: 99%

PGE₂-mediated cytoprotection in renal epithelial cells: evidence for a pharmacologically distinct receptor

Weber

Monks

Lau

1997

American Journal of Physiology-Renal Physiology

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Although the exact mechanism of prostaglandin E2(PGE2)-mediated cytoprotection has not been elucidated, its ability to induce cytoprotection in cell culture suggests this action occurs at the cellular level. The present studies were conducted to determine whether PGE2 induces protection against 2,3,5-(trisglutathion- S-yl)-hydroquinone [2,3,5-(trisglutathion- S-yl)-HQ]-mediated cytotoxicity in a renal proximal tubule epithelial cell line (LLC-PK1) and to delineate the cellular and molecular mechanisms associated with this response. Pretreatment of LLC-PK1 cells with 0.01–40 μM PGE2 for 24 h fully protects against a moderately toxic concentration of 2,3,5-(trisglutathion- S-yl)-HQ. PGE2-mediated cytoprotection is observed in cells pretreated at pH 7.4 but not at pH 7.8. However, cytoprotection is observed in LLC-PK1 cells pretreated with the PGE2 analog, 11-deoxy-16,16-dimethyl PGE2(DDM-PGE2) but not with the PGE2 receptor [E-prostanoid (EP)] agonists 17-phenyltrinor PGE2(EP1), 11-deoxy PGE1(EP2/EP4), sulprostone (EP1/EP3), PGE1, or PGA2. 12- O-tetradecanoylphorbol-13-acetate (TPA), a potent activator of protein kinase C (PKC), also induces cytoprotection, supporting a role for this pathway in the cytoprotective response. PGE2, DDM-PGE2, and TPA all induce the binding of nuclear proteins to a TPA responsive element (TRE), whereas analogs that did not induce cytoprotection (PGE1, 17-phenyltrinor PGE2, sulprostone) were without effect. DDM-PGE2- and TPA-mediated cytoprotection and TRE binding activity are inhibited by N-(2{[3-(4-bromophenyl)-2-propenyl]-amino}-ethyl)-5-isoquinolinesulfonamide (H-89), a PKC inhibitor. These data suggest that cytoprotection by PGE2 and DDM-PGE2 in LLC-PK1 cells is mediated by a PKC-coupled receptor, which is pharmacologically distinct from the presently classified EP receptor subtypes.

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Induction of Highly Proliferative Phenotypes in Cultured Glomerular Mesangial Cells by Benzo[a]pyrene Alone or in Combination with Methoxamine

Cited by 13 publications

References 0 publications

Acrolein Causes Inhibitor κB-independent Decreases in Nuclear Factor κB Activation in Human Lung Adenocarcinoma (A549) Cells

Acrolein Causes Inhibitor κB-independent Decreases in Nuclear Factor κB Activation in Human Lung Adenocarcinoma (A549) Cells

Hepatocyte growth factor induces tubulogenesis of primary renal proximal tubular epithelial cells

PGE₂-mediated cytoprotection in renal epithelial cells: evidence for a pharmacologically distinct receptor

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Induction of Highly Proliferative Phenotypes in Cultured Glomerular Mesangial Cells by Benzo[a]pyrene Alone or in Combination with Methoxamine

Cited by 13 publications

References 0 publications

Acrolein Causes Inhibitor κB-independent Decreases in Nuclear Factor κB Activation in Human Lung Adenocarcinoma (A549) Cells

Acrolein Causes Inhibitor κB-independent Decreases in Nuclear Factor κB Activation in Human Lung Adenocarcinoma (A549) Cells

Hepatocyte growth factor induces tubulogenesis of primary renal proximal tubular epithelial cells

PGE2-mediated cytoprotection in renal epithelial cells: evidence for a pharmacologically distinct receptor

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PGE₂-mediated cytoprotection in renal epithelial cells: evidence for a pharmacologically distinct receptor