Alleviation of apoptosis by serum in Chinese hamster ovary cells ectopically expressing human Fas antigen.
Fas-mediated apoptosis is an important regulatory mechanism for the development of T-cells and prevention of oncogenesis. Here, we establish Chinese hamster ovary (CHO) cell lines which stably express Fas antigen, and analyzed apoptosis induced by anti-Fas IgM. While Fas-transfected hamster cells did not undergo apoptosis when stimulated with anti-Fas antibody in the presence of medium containing 10% serum, in reduced serum concentrations, anti-Fas antibody caused these cells to round up and detach from the culture dish. Analysis of the DNA content by a flow cytometry demonstrated a significant increase of cells with sub-G1 amount of DNA upon Fas stimulation in the low serum concentrations. The increase in the number of apoptosis cells was inhibited by an apopain (CPP32, caspase 3) inhibitor or insulin-like growth factor-I. In contrast, apoptosis in a Fas-transfected mouse T-cell line occurred in the presence of 10% serum. these results suggest that factors including insulin-like growth factor-I in fetal bovine serum protect CHO cells from apopain-dependent apoptosis mediated by Fas-antigen stimulation.
Lee
Y S
YS
National Institute of Bioscience and Human-Technology, Tsukuba, Japan.
Nakajima
H
H
Chang
Y C
YC
Park
K I
KI
Mitsui
Y
Y
Magae
J
J
Saida
K
K
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology*; D016466:CHO Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D053148:Caspase 3; D020169:Caspases; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D006224:Cricetinae; D016895:Culture Media, Serum-Free / Q000737:chemistry / Q000494:pharmacology*; D003546:Cysteine Endopeptidases / Q000187:drug effects; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D053938:DNA Fragmentation / Q000187:drug effects; D005312:Fetal Blood / Q000737:chemistry; D005434:Flow Cytometry; D015870:Gene Expression / Q000235:genetics; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D009842:Oligopeptides / Q000494:pharmacology; D011994:Recombinant Proteins / Q000235:genetics; D019014:fas Receptor / Q000235:genetics* / Q000276:immunology / Q000502:physiology
D000888:Antibodies, Anti-Idiotypic; D016895:Culture Media, Serum-Free; D015853:Cysteine Proteinase Inhibitors; D009842:Oligopeptides; D011994:Recombinant Proteins; C044203:anti-IgM; C110772:benzoylcarbonyl-aspartyl-glutamyl-valyl-aspartyl-fluoromethyl ketone; D019014:fas Receptor; D007334:Insulin-Like Growth Factor I; C505407:CASP3 protein, human; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases