NMDA and HIV-1 coat protein, GP120, produce necrotic but not apoptotic cell death in human CHP100 neuroblastoma cultures via a mechanism involving calpain.
Treatment of neuroblastoma cultures with N-methyl-D-aspartate (NMDA) or human immunodeficiency virus type 1 (HIV-1) coat protein, gp120, induces significant cytotoxic effects which are reduced by leupeptin, E-64, N-Ac-Leu-Leu-norleucinal (ALLnL) as well as by N-Ac-Leu-Leu-normethioninal (ALLnM) and this suggests that activation of the Ca(2+)-dependent protease, calpain, is involved. The cell death induced by NMDA and gp120 appears to be of the necrotic type; in fact, analysis of DNA fragmentation by flow cytometry or agarose gel electrophoresis failed to demonstrate signs of apoptosis, such as the presence of apoptotic bodies or internucleosomal cleavage. Similar negative results were also obtained by studying the nuclear morphology of the cells with Hoechst 33258 staining. Altogether the data indicate that neuroblastoma cell death induced by NMDA and gp120 is of the necrotic type and this implicates calpain protease.
Corasaniti
M T
MT
Faculty of Pharmacy, University of Reggio Calabria, Catanzaro, Italy.
Navarra
M
M
Catani
M V
MV
Melino
G
G
Nisticò
G
G
Finazzi-Agrò
A
A
D002118:Calcium / Q000378:metabolism; D002154:Calpain / Q000037:antagonists & inhibitors / Q000378:metabolism*; D016923:Cell Death; D002467:Cell Nucleus / Q000648:ultrastructure; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004789:Enzyme Activation; D015699:HIV Envelope Protein gp120 / Q000633:toxicity*; D006801:Humans; D016202:N-Methylaspartate / Q000633:toxicity*; D009336:Necrosis; D009447:Neuroblastoma; D009474:Neurons / Q000187:drug effects / Q000201:enzymology / Q000473:pathology*; D009498:Neurotoxins / Q000633:toxicity*; D016194:Receptors, N-Methyl-D-Aspartate / Q000378:metabolism; D014407:Tumor Cells, Cultured
D015853:Cysteine Proteinase Inhibitors; D015699:HIV Envelope Protein gp120; D009498:Neurotoxins; D016194:Receptors, N-Methyl-D-Aspartate; D016202:N-Methylaspartate; D002154:Calpain; D002118:Calcium