Acid Ceramidase in Melanoma: EXPRESSION, LOCALIZATION, AND EFFECTS OF PHARMACOLOGICAL INHIBITION

Natalia Realini; Francesca Palese; Daniela Pizzirani; Silvia Pontis; Abdul Basit; Anders Bach; Anand Ganesan; Daniele Piomelli

doi:10.1074/jbc.M115.666909

Acid Ceramidase in Melanoma: EXPRESSION, LOCALIZATION, AND EFFECTS OF PHARMACOLOGICAL INHIBITION

Natalia Realini, Francesca Palese, Daniela Pizzirani, Silvia Pontis, Abdul Basit, Anders Bach, Anand Ganesan, Daniele Piomelli

47 Citations (Scopus)

Abstract

Acid ceramidase (AC) is a lysosomal cysteine amidase that controls sphingolipid signaling by lowering the levels of ceramides and concomitantly increasing those of sphingosine and its bioactive metabolite, sphingosine 1-phosphate. In the present study, we evaluated the role of AC-regulated sphingolipid signaling in melanoma. We found that AC expression is markedly elevated in normal human melanocytes and proliferative melanoma cell lines, compared with other skin cells (keratinocytes and fibroblasts) and non-melanoma cancer cells. High AC expression was also observed in biopsies from human subjects with Stage II melanoma. Immunofluorescence studies revealed that the subcellular localization of AC differs between melanocytes (where it is found in both cytosol and nucleus) and melanoma cells (where it is primarily localized to cytosol). In addition to having high AC levels, melanoma cells generate lower amounts of ceramides than normal melanocytes do. This down-regulation in ceramide production appears to result from suppression of the de novo biosynthesis pathway. To test whether AC might contribute to melanoma cell proliferation, we blocked AC activity using a new potent (IC50 = 12 nM) and stable inhibitor. AC inhibition increased cellular ceramide levels, decreased sphingosine 1-phosphate levels, and acted synergistically with several, albeit not all, antitumoral agents. The results suggest that AC-controlled sphingolipid metabolism may play an important role in the control of melanoma proliferation.

Original language	English
Journal	The Journal of Biological Chemistry
Volume	291
Issue number	5
Pages (from-to)	2422-34
Number of pages	13
ISSN	0021-9258
DOIs	https://doi.org/10.1074/jbc.M115.666909
Publication status	Published - 29 Jan 2016

Keywords

Acid Ceramidase
Cell Line, Tumor
Cell Proliferation
Cell Survival
Ceramides
Down-Regulation
Enzyme Inhibitors
Fibroblasts
Gene Expression Regulation, Neoplastic
HCT116 Cells
Hep G2 Cells
Humans
Inhibitory Concentration 50
Keratinocytes
Lipids
Lysophospholipids
MCF-7 Cells
Melanocytes
Melanoma
Microscopy, Confocal
Microscopy, Fluorescence
Oxidoreductases
RNA, Small Interfering
Serine C-Palmitoyltransferase
Signal Transduction
Skin Neoplasms
Sphingolipids
Sphingosine
Uracil
Journal Article

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1074/jbc.M115.666909

Cite this

@article{5e40bac7d82945a18bcf5f9bcd23f9bd,

title = "Acid Ceramidase in Melanoma: EXPRESSION, LOCALIZATION, AND EFFECTS OF PHARMACOLOGICAL INHIBITION",

abstract = "Acid ceramidase (AC) is a lysosomal cysteine amidase that controls sphingolipid signaling by lowering the levels of ceramides and concomitantly increasing those of sphingosine and its bioactive metabolite, sphingosine 1-phosphate. In the present study, we evaluated the role of AC-regulated sphingolipid signaling in melanoma. We found that AC expression is markedly elevated in normal human melanocytes and proliferative melanoma cell lines, compared with other skin cells (keratinocytes and fibroblasts) and non-melanoma cancer cells. High AC expression was also observed in biopsies from human subjects with Stage II melanoma. Immunofluorescence studies revealed that the subcellular localization of AC differs between melanocytes (where it is found in both cytosol and nucleus) and melanoma cells (where it is primarily localized to cytosol). In addition to having high AC levels, melanoma cells generate lower amounts of ceramides than normal melanocytes do. This down-regulation in ceramide production appears to result from suppression of the de novo biosynthesis pathway. To test whether AC might contribute to melanoma cell proliferation, we blocked AC activity using a new potent (IC50 = 12 nM) and stable inhibitor. AC inhibition increased cellular ceramide levels, decreased sphingosine 1-phosphate levels, and acted synergistically with several, albeit not all, antitumoral agents. The results suggest that AC-controlled sphingolipid metabolism may play an important role in the control of melanoma proliferation.",

keywords = "Acid Ceramidase, Cell Line, Tumor, Cell Proliferation, Cell Survival, Ceramides, Down-Regulation, Enzyme Inhibitors, Fibroblasts, Gene Expression Regulation, Neoplastic, HCT116 Cells, Hep G2 Cells, Humans, Inhibitory Concentration 50, Keratinocytes, Lipids, Lysophospholipids, MCF-7 Cells, Melanocytes, Melanoma, Microscopy, Confocal, Microscopy, Fluorescence, Oxidoreductases, RNA, Small Interfering, Serine C-Palmitoyltransferase, Signal Transduction, Skin Neoplasms, Sphingolipids, Sphingosine, Uracil, Journal Article",

author = "Natalia Realini and Francesca Palese and Daniela Pizzirani and Silvia Pontis and Abdul Basit and Anders Bach and Anand Ganesan and Daniele Piomelli",

note = "{\textcopyright} 2016 by The American Society for Biochemistry and Molecular Biology, Inc.",

year = "2016",

month = jan,

day = "29",

doi = "10.1074/jbc.M115.666909",

language = "English",

volume = "291",

pages = "2422--34",

journal = "The Journal of Biological Chemistry",

issn = "0021-9258",

publisher = "American Society for Biochemistry and Molecular Biology, Inc.",

number = "5",

}

TY - JOUR

T1 - Acid Ceramidase in Melanoma

T2 - EXPRESSION, LOCALIZATION, AND EFFECTS OF PHARMACOLOGICAL INHIBITION

AU - Realini, Natalia

AU - Palese, Francesca

AU - Pizzirani, Daniela

AU - Pontis, Silvia

AU - Basit, Abdul

AU - Bach, Anders

AU - Ganesan, Anand

AU - Piomelli, Daniele

PY - 2016/1/29

Y1 - 2016/1/29

N2 - Acid ceramidase (AC) is a lysosomal cysteine amidase that controls sphingolipid signaling by lowering the levels of ceramides and concomitantly increasing those of sphingosine and its bioactive metabolite, sphingosine 1-phosphate. In the present study, we evaluated the role of AC-regulated sphingolipid signaling in melanoma. We found that AC expression is markedly elevated in normal human melanocytes and proliferative melanoma cell lines, compared with other skin cells (keratinocytes and fibroblasts) and non-melanoma cancer cells. High AC expression was also observed in biopsies from human subjects with Stage II melanoma. Immunofluorescence studies revealed that the subcellular localization of AC differs between melanocytes (where it is found in both cytosol and nucleus) and melanoma cells (where it is primarily localized to cytosol). In addition to having high AC levels, melanoma cells generate lower amounts of ceramides than normal melanocytes do. This down-regulation in ceramide production appears to result from suppression of the de novo biosynthesis pathway. To test whether AC might contribute to melanoma cell proliferation, we blocked AC activity using a new potent (IC50 = 12 nM) and stable inhibitor. AC inhibition increased cellular ceramide levels, decreased sphingosine 1-phosphate levels, and acted synergistically with several, albeit not all, antitumoral agents. The results suggest that AC-controlled sphingolipid metabolism may play an important role in the control of melanoma proliferation.

AB - Acid ceramidase (AC) is a lysosomal cysteine amidase that controls sphingolipid signaling by lowering the levels of ceramides and concomitantly increasing those of sphingosine and its bioactive metabolite, sphingosine 1-phosphate. In the present study, we evaluated the role of AC-regulated sphingolipid signaling in melanoma. We found that AC expression is markedly elevated in normal human melanocytes and proliferative melanoma cell lines, compared with other skin cells (keratinocytes and fibroblasts) and non-melanoma cancer cells. High AC expression was also observed in biopsies from human subjects with Stage II melanoma. Immunofluorescence studies revealed that the subcellular localization of AC differs between melanocytes (where it is found in both cytosol and nucleus) and melanoma cells (where it is primarily localized to cytosol). In addition to having high AC levels, melanoma cells generate lower amounts of ceramides than normal melanocytes do. This down-regulation in ceramide production appears to result from suppression of the de novo biosynthesis pathway. To test whether AC might contribute to melanoma cell proliferation, we blocked AC activity using a new potent (IC50 = 12 nM) and stable inhibitor. AC inhibition increased cellular ceramide levels, decreased sphingosine 1-phosphate levels, and acted synergistically with several, albeit not all, antitumoral agents. The results suggest that AC-controlled sphingolipid metabolism may play an important role in the control of melanoma proliferation.

KW - Acid Ceramidase

KW - Cell Line, Tumor

KW - Cell Proliferation

KW - Cell Survival

KW - Ceramides

KW - Down-Regulation

KW - Enzyme Inhibitors

KW - Fibroblasts

KW - Gene Expression Regulation, Neoplastic

KW - HCT116 Cells

KW - Hep G2 Cells

KW - Humans

KW - Inhibitory Concentration 50

KW - Keratinocytes

KW - Lipids

KW - Lysophospholipids

KW - MCF-7 Cells

KW - Melanocytes

KW - Melanoma

KW - Microscopy, Confocal

KW - Microscopy, Fluorescence

KW - Oxidoreductases

KW - RNA, Small Interfering

KW - Serine C-Palmitoyltransferase

KW - Signal Transduction

KW - Skin Neoplasms

KW - Sphingolipids

KW - Sphingosine

KW - Uracil

KW - Journal Article

U2 - 10.1074/jbc.M115.666909

DO - 10.1074/jbc.M115.666909

M3 - Journal article

C2 - 26553872

SN - 0021-9258

VL - 291

SP - 2422

EP - 2434

JO - The Journal of Biological Chemistry

JF - The Journal of Biological Chemistry

IS - 5

ER -

Acid Ceramidase in Melanoma: EXPRESSION, LOCALIZATION, AND EFFECTS OF PHARMACOLOGICAL INHIBITION

Abstract

Keywords

UN SDGs

Access to Document

Fingerprint

Cite this