The mechanism of thioacetamide-induced apoptosis in the L37 albumin-SV40 T-antigen transgenic rat hepatocyte-derived cell line occurs without DNA fragmentation.
The hepatotoxicant thioacetamide (TH) has classically been used as a model to study hepatic necrosis; however, recent studies have shown that TH can also induce apoptosis. In this report we demonstrate that 2.68+/-0.54% of the albumin-SV40 T-antigen transgenic rat hepatocytes undergo TH-induced apoptosis, a level comparable to other in vivo models of liver apoptosis. In addition, TH could induce apoptosis and necrosis in the L37 albumin-SV40 T-antigen transgenic rat liver-derived cell line. Examination of dying L37 cells treated with 100 mM TH by electron microscopy revealed distinct morphological characteristics that could be attributed to apoptosis. Quantitation of apoptosis by FACS analysis 24 h after treatment with 100 mM TH revealed that 81.3+/-1.6% of the cells were undergoing apoptosis. In contrast, when L37 cells were treated with 250 mM TH, cells exhibited characteristics consistent with necrotic cell death. DNA fragmentation ladders were produced by growth factor withdrawal-induced apoptosis; however, in 100 mM TH-induced apoptosis, DNA fragmentation ladders were not observed. Analysis of endonuclease activity in L37 cells revealed that the enzymes were not inactivated in the presence of 100 mM TH. The data presented in this report indicate that the L37 cell line could be used to study the mechanism of TH-induced apoptosis that was not mediated through a mechanism requiring DNA fragmentation.
Bulera
S J
SJ
Department of Pathology and Experimental Toxicology, Parke-Davis/Warner-Lambert Company, Ann Arbor, Michigan 48105, USA.
Sattler
C A
CA
Gast
W L
WL
Heath
S
S
Festerling
T A
TA
Pitot
H C
HC
D000418:Albumins / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics*; D017209:Apoptosis / Q000187:drug effects*; D002460:Cell Line; D002469:Cell Separation; D003851:Deoxyribonucleases / Q000378:metabolism; D005434:Flow Cytometry; D008099:Liver / Q000166:cytology* / Q000201:enzymology; D051381:Rats; D013539:Simian virus 40 / Q000276:immunology; D013853:Thioacetamide / Q000494:pharmacology*
D000418:Albumins; D000952:Antigens, Polyomavirus Transforming; D013853:Thioacetamide; D003851:Deoxyribonucleases