Palmitoyl-ceramide accumulation with necrotic cell death in A549 cells, followed by a steep increase in sphinganine content
| Autor: | Mototeru Yamane |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2015 |
| Předmět: |
Necrosis
Research paper acridine orange 3 6-Bis(dimethylamino) acridine hydrochloride CathB cathepsin B 4-HPR N-(4-hydroxyphenyl)retinamide ([13C16]C16:0-CoA palmitoyl-13C16 coenzyme A Biochemistry Cathepsin B chemistry.chemical_compound SIM selected-ion monitoring C2:0-CoA acetyl-coenzyme A DL-PDMP DL-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol lcsh:QD415-436 A549 cells Ser Serine IS internal standard CHOP CAAT/enhancer binding protein homologous protein DMEM Dulbecco's modified Eagle's medium ESI electrospray ionization LDH lactate dehydrogenase [1 2 3 4-13C4]C16:0 acid palmitic acid-1 2 3 4-13C4 APCI atmospheric pressure chemical ionization SDS-PAGE sodium dodecyl sulfate-polyacrylamide gel electrophoresis [2-13C]C2:0 acid sodium acetate-2-13C SPTLC SPT-long chain base subunit d18:1-[D31]C16:0-Cer N-palmitoyl [D31]-D-erythro-sphingosine Lys lysosomes Ceramidase Cell biology lipids (amino acids peptides and proteins) medicine.symptom D-NMAPPD D-NMAPPD N-[(1R 2R)-2-hydroxy-1-(hydroxy-methyl)-2-(4-nitrophenyl)ethyl]tetradecanamide Ceramide C16:0-CoA palmitoyl-coenzyme A DMSO dimethylsulfoxide SDS sodium dodecyl sulfate GlcCer glucosylceramide Palmitoyl-ceramide Cer ceramide [D7]d18:1 D-erythro-sphingosine-D7 Biology Myriocin 2-amino-3 4-dihydroxy-2-(hydroxymethyl)-14-oxo-6-eicosenoic acid C16:0-Cer palmitoyl-ceramide lcsh:Biochemistry ER endoplasmic reticulum L-[2 3 3-D3]Ser L-serine-2 3 3-D3 FBS fetal bovine serum Survivin medicine d18:1 sphingosine DL-PDMP LC3 microtubule-associated protein 1 light chain 3B A549 cell SM sphingomyelin Cell growth SPT serine palmitoyltransferase [D7]d18:0 D-erythro-sphinganine-D7 LMP lysosomal membrane permeabilization FATP1 fatty acid transport protein 1 carbohydrates (lipids) Sphinganine chemistry Apoptosis DTT dithiothreitol CerS ceramide synthase BSA bovine serum albumin d18:0 sphinganine MAM mitochondria-associated membrane DAPI 4′ 6-diamidino-2-phenylindole |
| Zdroj: | Biochimie Open Biochimie Open, Vol 1, Iss C, Pp 11-27 (2015) |
| ISSN: | 2214-0085 |
| Popis: | Ceramides (Cers) have recently been identified as key signaling molecules that mediate biological functions such as cell growth, differentiation, senescence, apoptosis, and autophagy. However, the functions of Cer accumulation in necrotic cell death remain unknown. The aim of this study was to clarify the relationship between Cer accumulation with inhibition of the conversion pathway of Cer and concomitant necrotic cell death. In order to minimize the effect of apoptosis against necrotic cell death, A549 cells having the inhibiting effect of caspase 9 by survivin were used in this study. Consequently, Cer accumulation in A549 cells would likely be associated with a pathway other than the mitochondrial caspase-dependent pathway of apoptosis. Here, we showed that the dual addition of a glucosyl-Cer synthase inhibitor and a ceramidase inhibitor to A549 cell culture induced palmitoyl-Cer accumulation with Cer synthase 5 expression and necrotic cell death with lysosomal rupture together with leakage of cathepsin B/alkalization after 2–3 h, although it is unknown in this study whether the necrotic cell death was caused by the lysosomal rupture. This Cer accumulation was followed by a steep increase in sphinganine base levels via the activation of serine palmitoyltransferase activity brought about by the increase in palmitoyl-coenzyme A concentration as a substrate after 5–6 h. The increase in palmitoyl-coenzyme A concentration was achieved by activation of the fatty acid synthetic pathway from acetyl coenzyme A. Highlights • Palmitoylceramide accumulation with necrosis is studied. • The accumulation is with ceramide synthase 5 expression. • The accumulation is followed by high sphinganine levels. |
| Databáze: | OpenAIRE |
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