Alterations of extracellular calcium elicit selective modes of cell death and protease activation in SH-SY5Y human neuroblastoma cells

被引:77
作者
McGinnis, KM
Wang, KKW
Gnegy, ME
机构
[1] Univ Michigan, Sch Med, Dept Pharmacol, Ann Arbor, MI 48109 USA
[2] Warner Lambert Co, Parke Davis Pharmaceut Res Div, Dept Neurosci Therapeut, Lab Neurobiochem, Ann Arbor, MI USA
关键词
apoptosis; necrosis; calpain; caspase-3; Ca2+ homeostasis; neuronal death;
D O I
10.1046/j.1471-4159.1999.0721853.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The role of intracellular Ca2+ homeostasis in mechanisms of neuronal cell death and cysteine protease activation was investigated in SH-SY5Y human neuroblastoma cells. Cells were incubated in 2 mM EGTA to lower intracellular Ca2+ or 5 mM CaCl2 to raise it. Cell death and activation of calpain and caspase-3 were measured. Both EGTA and excess CaCl2 elicited cell death. EGTA induced DNA laddering and an increase in caspase-3-like, but not calpain, activity. Pan-caspase inhibitors protected against EGTA-, but not CaCl2-, induced cell death. Conversely, excess Ca2+ elicited necrosis and activated calpain but not caspase-3. Calpain inhibitors did not preserve cell viability. Ca2+ was the death-mediating factor, because restoration of extracellular Ca2+ protected against cell death induced by EGTA and blockade of Ca2+ channels by Ni2+ protected against that induced by high Ca2+. We conclude that the EGTA treatment lowered intracellular Ca2+ and elicited caspase-3-like protease activity, which led to apoptosis. Conversely, excess extracellular Ca2+ entered Ca2+ channels and increased intracellular Ca2+ leading to calpain activation and necrosis, The mode of cell death and protease activation in response to changing Ca2+ were selective and mutually exclusive, demonstrating that these are useful models to individually investigate apoptosis and necrosis.
引用
收藏
页码:1853 / 1863
页数:11
相关论文
共 56 条
[21]   PROGRAMMED CELL-DEATH - ITS POSSIBLE CONTRIBUTION TO NEUROTOXICITY MEDIATED BY CALCIUM-CHANNEL ANTAGONISTS [J].
KOH, JY ;
COTMAN, CW .
BRAIN RESEARCH, 1992, 587 (02) :233-240
[22]   CLEAVAGE OF POLY(ADP-RIBOSE) POLYMERASE BY A PROTEINASE WITH PROPERTIES LIKE ICE [J].
LAZEBNIK, YA ;
KAUFMANN, SH ;
DESNOYERS, S ;
POIRIER, GG ;
EARNSHAW, WC .
NATURE, 1994, 371 (6495) :346-347
[23]  
MALAYEV A, 1995, J MEMBRANE BIOL, V146, P101
[24]   PROTEOLYSIS OF FODRIN (NONERYTHROID SPECTRIN) DURING APOPTOSIS [J].
MARTIN, SJ ;
OBRIEN, GA ;
NISHIOKA, WK ;
MCGAHON, AJ ;
MAHBOUBI, A ;
SAIDO, TC ;
GREEN, DR .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (12) :6425-6428
[25]  
Mattson M P, 1990, Adv Exp Med Biol, V268, P211
[26]   The role of calcium in the regulation of apoptosis [J].
McConkey, DJ ;
Orrenius, S .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 1997, 239 (02) :357-366
[27]   GLUCOCORTICOIDS ACTIVATE A SUICIDE PROCESS IN THYMOCYTES THROUGH AN ELEVATION OF CYTOSOLIC CA-2+ CONCENTRATION [J].
MCCONKEY, DJ ;
NICOTERA, P ;
HARTZELL, P ;
BELLOMO, G ;
WYLLIE, AH ;
ORRENIUS, S .
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, 1989, 269 (01) :365-370
[28]  
McGinnis KM, 1998, J NEUROCHEM, V70, P139
[29]   Altered bcl-2 and bax expression and intracellular Ca2+ signaling in apoptosis of pancreatic cells and the impairment of glucose-induced insulin secretion [J].
Mizuno, N ;
Yoshitomi, H ;
Ishida, H ;
Kuromi, H ;
Kawaki, J ;
Seino, Y ;
Seino, S .
ENDOCRINOLOGY, 1998, 139 (03) :1429-1439
[30]  
Munir M, 1995, J NEUROSCI, V15, P7847