SGK-1 protects kidney cells against apoptosis induced by ceramide and TNF-α

D. Pastore, David Della Morte, A. Coppola, B. Capuani, M. F. Lombardo, F. Pacifici, F. Ferrelli, R. Arriga, C. Mammi, M. Federici, A. Bellia, N. Di Daniele, M. Tesauro, G. Donadel, D. Noto, P. Sbraccia, G. Sconocchia, D. Lauro

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Ceramide regulates several different cellular responses including mechanisms leading to apoptosis. Serum-and glucocorticoidinducible protein kinase (SGK)-1 is a serine threonine kinase, which activates survival pathways in response to stress stimuli. Recently, we demonstrated an anti-apoptotic role of SGK-1 in human umbilical endothelial cells treated with high glucose. In the present study, since ceramide induces apoptosis by multiple mechanisms in diabetes and its complication such as nephropathy, we aimed to investigate whether SGK-1 may protect even against apoptosis induced by ceramide in kidney cells. Human embryonic kidney (HEK)-293 cells stable transfected with SGK-1 wild type (SGK-1wt) and its dominant negative gene (SGK-1dn) have been used in this study. Apoptotic stimuli were induced by C2-ceramide and TNF-α to increase endogenous synthesis of ceramide. Upon activation with these stimuli, SGK-1wt transfected cells have a statistically significant reduction of apoptosis compared with SGK-1dn cells (P<0.001). This protection was dependent on activation of caspase-3 and Poly-ADP-ribosepolymerase-1 (PARP-1) cleavage. SGK-1 and AKT-1 two highly homologous kinases differently reacted to ceramide treatment, since SGK-1 increases in response to apoptotic stimulus while AKT-1 decreases. This enhancement of SGK-1 was dependent on p38-mitogen-activated-protein kinases (p38MAPK), cyclic-adenosine-monophosphate/protein kinase A (cAMP/PKA) and phosphoinositide-3-kinase (PI3K) pathways. Especially, by using selective LY294002 inhibitor, we demonstrated that the most involved pathway in the SGK-1 mediated process of protection was PI3K. Treatment with inhibitor of SGK-1 (GSK650394) significantly enhanced TNF-α-dependent apoptosis in HEK-293 cells overexpressing SGK-1wt. Caspase-3,-8 and-9 selective inhibitors confirmed that SGK-1 reduced the activation of caspase-dependent apoptosis, probably by both intrinsic and extrinsic pathways. In conclusion, we demonstrated that in kidney cells, overexpression of SGK-1 is protective against ceramide-induced apoptosis and the role of SGK-1 can be potentially explored as a therapeutic target in conditions like diabetes, where ceramide levels are increased.

Original languageEnglish (US)
Article numbere1890
JournalCell Death and Disease
Volume6
Issue number9
DOIs
StatePublished - Sep 17 2016
Externally publishedYes

Fingerprint

Ceramides
Protein Kinases
Blood Proteins
Apoptosis
Kidney
1-Phosphatidylinositol 4-Kinase
Caspase 3
Umbilicus
Dominant Genes
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
Caspase 8
Protein-Serine-Threonine Kinases
p38 Mitogen-Activated Protein Kinases
Diabetes Complications
Caspases
Cyclic AMP-Dependent Protein Kinases
Cyclic AMP
Adenosine Diphosphate

ASJC Scopus subject areas

  • Immunology
  • Cellular and Molecular Neuroscience
  • Cell Biology
  • Cancer Research

Cite this

Pastore, D., Della Morte, D., Coppola, A., Capuani, B., Lombardo, M. F., Pacifici, F., ... Lauro, D. (2016). SGK-1 protects kidney cells against apoptosis induced by ceramide and TNF-α. Cell Death and Disease, 6(9), [e1890]. https://doi.org/10.1038/cddis.2015.232

SGK-1 protects kidney cells against apoptosis induced by ceramide and TNF-α. / Pastore, D.; Della Morte, David; Coppola, A.; Capuani, B.; Lombardo, M. F.; Pacifici, F.; Ferrelli, F.; Arriga, R.; Mammi, C.; Federici, M.; Bellia, A.; Di Daniele, N.; Tesauro, M.; Donadel, G.; Noto, D.; Sbraccia, P.; Sconocchia, G.; Lauro, D.

In: Cell Death and Disease, Vol. 6, No. 9, e1890, 17.09.2016.

Research output: Contribution to journalArticle

Pastore, D, Della Morte, D, Coppola, A, Capuani, B, Lombardo, MF, Pacifici, F, Ferrelli, F, Arriga, R, Mammi, C, Federici, M, Bellia, A, Di Daniele, N, Tesauro, M, Donadel, G, Noto, D, Sbraccia, P, Sconocchia, G & Lauro, D 2016, 'SGK-1 protects kidney cells against apoptosis induced by ceramide and TNF-α', Cell Death and Disease, vol. 6, no. 9, e1890. https://doi.org/10.1038/cddis.2015.232
Pastore, D. ; Della Morte, David ; Coppola, A. ; Capuani, B. ; Lombardo, M. F. ; Pacifici, F. ; Ferrelli, F. ; Arriga, R. ; Mammi, C. ; Federici, M. ; Bellia, A. ; Di Daniele, N. ; Tesauro, M. ; Donadel, G. ; Noto, D. ; Sbraccia, P. ; Sconocchia, G. ; Lauro, D. / SGK-1 protects kidney cells against apoptosis induced by ceramide and TNF-α. In: Cell Death and Disease. 2016 ; Vol. 6, No. 9.
@article{572b591e097546bd8de42795420f5fb2,
title = "SGK-1 protects kidney cells against apoptosis induced by ceramide and TNF-α",
abstract = "Ceramide regulates several different cellular responses including mechanisms leading to apoptosis. Serum-and glucocorticoidinducible protein kinase (SGK)-1 is a serine threonine kinase, which activates survival pathways in response to stress stimuli. Recently, we demonstrated an anti-apoptotic role of SGK-1 in human umbilical endothelial cells treated with high glucose. In the present study, since ceramide induces apoptosis by multiple mechanisms in diabetes and its complication such as nephropathy, we aimed to investigate whether SGK-1 may protect even against apoptosis induced by ceramide in kidney cells. Human embryonic kidney (HEK)-293 cells stable transfected with SGK-1 wild type (SGK-1wt) and its dominant negative gene (SGK-1dn) have been used in this study. Apoptotic stimuli were induced by C2-ceramide and TNF-α to increase endogenous synthesis of ceramide. Upon activation with these stimuli, SGK-1wt transfected cells have a statistically significant reduction of apoptosis compared with SGK-1dn cells (P<0.001). This protection was dependent on activation of caspase-3 and Poly-ADP-ribosepolymerase-1 (PARP-1) cleavage. SGK-1 and AKT-1 two highly homologous kinases differently reacted to ceramide treatment, since SGK-1 increases in response to apoptotic stimulus while AKT-1 decreases. This enhancement of SGK-1 was dependent on p38-mitogen-activated-protein kinases (p38MAPK), cyclic-adenosine-monophosphate/protein kinase A (cAMP/PKA) and phosphoinositide-3-kinase (PI3K) pathways. Especially, by using selective LY294002 inhibitor, we demonstrated that the most involved pathway in the SGK-1 mediated process of protection was PI3K. Treatment with inhibitor of SGK-1 (GSK650394) significantly enhanced TNF-α-dependent apoptosis in HEK-293 cells overexpressing SGK-1wt. Caspase-3,-8 and-9 selective inhibitors confirmed that SGK-1 reduced the activation of caspase-dependent apoptosis, probably by both intrinsic and extrinsic pathways. In conclusion, we demonstrated that in kidney cells, overexpression of SGK-1 is protective against ceramide-induced apoptosis and the role of SGK-1 can be potentially explored as a therapeutic target in conditions like diabetes, where ceramide levels are increased.",
author = "D. Pastore and {Della Morte}, David and A. Coppola and B. Capuani and Lombardo, {M. F.} and F. Pacifici and F. Ferrelli and R. Arriga and C. Mammi and M. Federici and A. Bellia and {Di Daniele}, N. and M. Tesauro and G. Donadel and D. Noto and P. Sbraccia and G. Sconocchia and D. Lauro",
year = "2016",
month = "9",
day = "17",
doi = "10.1038/cddis.2015.232",
language = "English (US)",
volume = "6",
journal = "Cell Death and Disease",
issn = "2041-4889",
publisher = "Nature Publishing Group",
number = "9",

}

TY - JOUR

T1 - SGK-1 protects kidney cells against apoptosis induced by ceramide and TNF-α

AU - Pastore, D.

AU - Della Morte, David

AU - Coppola, A.

AU - Capuani, B.

AU - Lombardo, M. F.

AU - Pacifici, F.

AU - Ferrelli, F.

AU - Arriga, R.

AU - Mammi, C.

AU - Federici, M.

AU - Bellia, A.

AU - Di Daniele, N.

AU - Tesauro, M.

AU - Donadel, G.

AU - Noto, D.

AU - Sbraccia, P.

AU - Sconocchia, G.

AU - Lauro, D.

PY - 2016/9/17

Y1 - 2016/9/17

N2 - Ceramide regulates several different cellular responses including mechanisms leading to apoptosis. Serum-and glucocorticoidinducible protein kinase (SGK)-1 is a serine threonine kinase, which activates survival pathways in response to stress stimuli. Recently, we demonstrated an anti-apoptotic role of SGK-1 in human umbilical endothelial cells treated with high glucose. In the present study, since ceramide induces apoptosis by multiple mechanisms in diabetes and its complication such as nephropathy, we aimed to investigate whether SGK-1 may protect even against apoptosis induced by ceramide in kidney cells. Human embryonic kidney (HEK)-293 cells stable transfected with SGK-1 wild type (SGK-1wt) and its dominant negative gene (SGK-1dn) have been used in this study. Apoptotic stimuli were induced by C2-ceramide and TNF-α to increase endogenous synthesis of ceramide. Upon activation with these stimuli, SGK-1wt transfected cells have a statistically significant reduction of apoptosis compared with SGK-1dn cells (P<0.001). This protection was dependent on activation of caspase-3 and Poly-ADP-ribosepolymerase-1 (PARP-1) cleavage. SGK-1 and AKT-1 two highly homologous kinases differently reacted to ceramide treatment, since SGK-1 increases in response to apoptotic stimulus while AKT-1 decreases. This enhancement of SGK-1 was dependent on p38-mitogen-activated-protein kinases (p38MAPK), cyclic-adenosine-monophosphate/protein kinase A (cAMP/PKA) and phosphoinositide-3-kinase (PI3K) pathways. Especially, by using selective LY294002 inhibitor, we demonstrated that the most involved pathway in the SGK-1 mediated process of protection was PI3K. Treatment with inhibitor of SGK-1 (GSK650394) significantly enhanced TNF-α-dependent apoptosis in HEK-293 cells overexpressing SGK-1wt. Caspase-3,-8 and-9 selective inhibitors confirmed that SGK-1 reduced the activation of caspase-dependent apoptosis, probably by both intrinsic and extrinsic pathways. In conclusion, we demonstrated that in kidney cells, overexpression of SGK-1 is protective against ceramide-induced apoptosis and the role of SGK-1 can be potentially explored as a therapeutic target in conditions like diabetes, where ceramide levels are increased.

AB - Ceramide regulates several different cellular responses including mechanisms leading to apoptosis. Serum-and glucocorticoidinducible protein kinase (SGK)-1 is a serine threonine kinase, which activates survival pathways in response to stress stimuli. Recently, we demonstrated an anti-apoptotic role of SGK-1 in human umbilical endothelial cells treated with high glucose. In the present study, since ceramide induces apoptosis by multiple mechanisms in diabetes and its complication such as nephropathy, we aimed to investigate whether SGK-1 may protect even against apoptosis induced by ceramide in kidney cells. Human embryonic kidney (HEK)-293 cells stable transfected with SGK-1 wild type (SGK-1wt) and its dominant negative gene (SGK-1dn) have been used in this study. Apoptotic stimuli were induced by C2-ceramide and TNF-α to increase endogenous synthesis of ceramide. Upon activation with these stimuli, SGK-1wt transfected cells have a statistically significant reduction of apoptosis compared with SGK-1dn cells (P<0.001). This protection was dependent on activation of caspase-3 and Poly-ADP-ribosepolymerase-1 (PARP-1) cleavage. SGK-1 and AKT-1 two highly homologous kinases differently reacted to ceramide treatment, since SGK-1 increases in response to apoptotic stimulus while AKT-1 decreases. This enhancement of SGK-1 was dependent on p38-mitogen-activated-protein kinases (p38MAPK), cyclic-adenosine-monophosphate/protein kinase A (cAMP/PKA) and phosphoinositide-3-kinase (PI3K) pathways. Especially, by using selective LY294002 inhibitor, we demonstrated that the most involved pathway in the SGK-1 mediated process of protection was PI3K. Treatment with inhibitor of SGK-1 (GSK650394) significantly enhanced TNF-α-dependent apoptosis in HEK-293 cells overexpressing SGK-1wt. Caspase-3,-8 and-9 selective inhibitors confirmed that SGK-1 reduced the activation of caspase-dependent apoptosis, probably by both intrinsic and extrinsic pathways. In conclusion, we demonstrated that in kidney cells, overexpression of SGK-1 is protective against ceramide-induced apoptosis and the role of SGK-1 can be potentially explored as a therapeutic target in conditions like diabetes, where ceramide levels are increased.

UR - http://www.scopus.com/inward/record.url?scp=84984699073&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84984699073&partnerID=8YFLogxK

U2 - 10.1038/cddis.2015.232

DO - 10.1038/cddis.2015.232

M3 - Article

C2 - 26379195

AN - SCOPUS:84984699073

VL - 6

JO - Cell Death and Disease

JF - Cell Death and Disease

SN - 2041-4889

IS - 9

M1 - e1890

ER -