Palmitoyl-ceramide accumulation with necrotic cell death in A549 cells, followed by a steep increase in sphinganine content

Yamane, Mototeru

doi:10.1016/j.biopen.2015.06.001

Cited by 4 publications

(16 citation statements)

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“…The protein concentration of the homogenate or the suspension of the lysosome-rich fraction was measured using BCA Protein Assay Reagents (Thermo Scientific). Extraction of Cers and dihydro-Cers in the homogenate or the suspension from the lysosome-rich fraction were performed using d18:1-[D 31 ]C16:0-Cer as the IS as described previously [7] . The extract was examined using an HPLC- atmospheric pressure chemical ionization (APCI)-mass spectrometry (MS) system.…”

Section: Methodsmentioning

confidence: 99%

“…Extraction of sphinganine (d18:0) and sphingosine (d18:1) in the homogenate described above or the suspension of the lysosome-rich fraction were performed using [D 7 ]d18:0 or [D 7 ]d18:1 as the IS as described previously [7] . The extract was examined using an HPLC-APCI-MS system.…”

Section: Methodsmentioning

confidence: 99%

“…To determine the d18:0, d18:1, Cer or dihydro-Cer content, HPLC-MS was performed using a Shimadzu (Kyoto, Japan) LCMS-2010EV mounted on an APCI probe, a quadrupole mass spectrometer, and connected reversed-phase HPLC separation as described previously [7] .…”

Section: Methodsmentioning

confidence: 99%

“…The quantitative determination of Cers/dihydro-Cers/d18:0/d18:1 by selected ion monitoring (SIM) using HPLC-APCI-MS was performed as described previously [7] .…”

Section: Methodsmentioning

confidence: 99%

“…Previously, we showed that a high concentration of DL-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol [DL-PDMP, an inhibitor of glucosyl(Glc)-Cer synthase] in A549 cell culture caused massive autophagy with endoplasmic reticulum stress and endogenous C16:0-Cer accumulation via Cer synthase (CerS) 5 protein expression in A549 cells, followed by autophagic cell death 24 h after treatment [6] . Furthermore, we showed that the dual addition of DL-PDMP and N-[(1R,2R)-2-hydroxy-1-(hydroxy-methyl)-2-(4-nitrophenyl)ethyl]tetradecanamide (D-NMAPPD, an inhibitor of ceramidase) to A549 cell culture induced additional endogenous C16:0-Cer accumulation with CerS5 expression and necrotic cell death with lysosomal rupture along with the leakage of cathepsin B/alkalization 2–3 h afterwards [7] . If C16:0-Cer channels were formed in the lysosome membrane with endogenous C16:0-Cer accumulation via the activation of CerS5 and the inhibition of lysosomal acid ceramidase by D-NMAPPD, this possibility would be of interest as a function of the liberation of cathepsin B from lysosomes causing necrotic cell death via C16:0-Cer channels in the lysosome membrane distinct from the mitochondrial caspase-dependent pathway of apoptosis.…”

Section: Introductionmentioning

confidence: 99%

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Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis

Yamane

Moriya

Kokuba

2017

Biochemistry and Biophysics Reports

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In this study, we showed that the dual addition of glucosyl ceramide synthase and ceramidase inhibitors to A549 cell culture led to the possibility of ceramide channel formation via endogenous palmitoyl-ceramide accumulation with an increase in cholesterol contents in the lysosome membrane as an initial step prior to initiation of necrotic cell death. In addition, the dual addition led to black circular structures of 10–20 nm, interpreted as stain-filled cylindrical channels on transmission electron microscopy. The formation of palmitoyl-ceramide channels in the lysosome membrane causes the liberation of cathepsin B from lysosomes for necrotic cell death. On the other hand, necrotic cell death in the dual addition was not caused by oxidative stress or cathepsin B activity, and the cell death was free from the contribution of the translation of Bax protein to the lysosome membrane.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…The quantitative determination of Cers/dihydro-Cers/d18:0/d18:1 by selected ion monitoring (SIM) using HPLC-APCI-MS was performed as described previously [7] .…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis

Yamane

Moriya

Kokuba

2017

Biochemistry and Biophysics Reports

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Evaluating Particle Emissions and Toxicity of 3D Pen Printed Filaments with Metal Nanoparticles As Additives: In Vitro and in Silico Discriminant Function Analysis

Singh

Maharjan

Jungnickel

et al. 2021

ACS Sustainable Chem. Eng.

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The emergence of low-cost 3D printing (particularly with 3D pens) in the home and school environment is a matter of concern since there is a lack of data confirming its safe application. The filament washed with water and the particle emission during printing (captured in water) were analyzed. In this report, we provide important insights about toxicity, intracellular mitochondrial stress, and cellular metabolic alterations as a mechanistic response after exposure of emissions released during 3D pen printing with polylactic acid (PLA) and PLA filaments with additives (carbon nanotubes (CNTs), copper, or steel). We adopted the design and exposure of a realistic inhalation procedure of aerosols via introducing mist-containing particles released during 3D pen printing processes of filaments with/without additive (CNTs, steel, copper) to A549 cells in a cloud chamber. The filaments with or without additives appear benign except for copper, which causes higher changes in stress, cell death, and metabolic perturbations. Albeit live/dead cell assay fluorescence-activated cell sorting analysis revealed little or no toxicity, time-of-flight secondary ion mass spectrometry bimolecular imaging and chemical mapping show insightful changes in cell membrane lipid compositions. Biochemical profiles obtained based on ion m/z signature peaks exhibit membrane and lipoprotein metabolism alterations which are similar to those adopted by A549 cells while developing drug resistance and their activation of the expression of their efflux pump. The results sought in this report suggest the cautious use of 3D pen printers for filaments embedded with redox active metals as additives in a home environment.

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Role of Glucocerebrosidase in Metabolic Reprogramming and Prognosis of Hepatocellular Carcinoma: Insights from a Comprehensive Gene Expression Analysis

Li,

Ma,

Wang

et al. 2024

Preprint

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Introduction: Hepatocellular carcinoma (HCC) poses a substantial global public health concern, with its intricate pathogenesis remaining incompletely elucidated. Metabolic reprogramming is pivotal in liver cancer progression. This study investigates the role of the lysosomal enzyme Glucocerebrosidase (GBA) in HCC initiation. Methods: We analyzed GBA-related gene expressions in 1003 primary liver cancer samples from the GEO database and 433 liver cancer samples from the TCGA database to examine GBA expression patterns and their association with liver cancer prognosis. Additionally, we manipulated GBA and glucosylceramide synthase (UGCG) expressions in the MHCC-97H cell line to investigate their effects on lysosomal and non-lysosomal metabolic genes. Results: GBA expression was significantly elevated in liver cancer samples and closely associated with poor prognosis. Overexpression of GBA led to upregulation of related lysosomal metabolic genes (NEU1, CTSD, CTSA, GALNS, GLB1) and non-lysosomal metabolic genes (ACOT8, FDPS, PMVK, PIGC, B4GALT3). Non-lysosomal genes were involved in N-acetyl metabolism, fatty acid β-oxidation, and cholesterol synthesis. Co-upregulation of UGCG and GBA resulted in a dose-dependent increase in ACOT family gene expressions (ACOT8, ACOT4, ACOT9, ACOT11). Survival analysis indicated high expression of these genes was related to lower short-term survival rates in liver cancer patients. Conclusion: Our findings suggest GBA plays a role in the metabolic reprogramming of HCC, influencing disease progression and prognosis by modulating genes involved in N-acetyl metabolism and lysosomal complexes. Downregulating GBA expression may present a potential therapeutic strategy for managing HCC.

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Palmitoyl-ceramide accumulation with necrotic cell death in A549 cells, followed by a steep increase in sphinganine content

Cited by 4 publications

References 34 publications

Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis

Visualization of ceramide channels in lysosomes following endogenous palmitoyl-ceramide accumulation as an initial step in the induction of necrosis

Evaluating Particle Emissions and Toxicity of 3D Pen Printed Filaments with Metal Nanoparticles As Additives: In Vitro and in Silico Discriminant Function Analysis

Role of Glucocerebrosidase in Metabolic Reprogramming and Prognosis of Hepatocellular Carcinoma: Insights from a Comprehensive Gene Expression Analysis

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