1. The Mechanism of Cell Death in Human Cultured Colon Adenocarcinoma Cell Line COLO 201 Induced by .BETA.-D-N-Acetylglucosaminyl-p-Nitrophenol
- Author
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Ikuko Kakizaki, Masahiko Endo, Akihiko Matsuki, Keiichi Takagaki, Akihiro Munakata, and Junko Kukidome
- Subjects
inorganic chemicals ,Programmed cell death ,Time Factors ,Cell Culture Techniques ,Apoptosis ,Adenocarcinoma ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Acetylglucosamine ,Nitrophenols ,Tumor Cells, Cultured ,medicine ,Humans ,heterocyclic compounds ,Dose-Response Relationship, Drug ,Molecular Structure ,General Medicine ,medicine.disease ,Molecular biology ,enzymes and coenzymes (carbohydrates) ,genomic DNA ,Biochemistry ,Cell culture ,Colonic Neoplasms ,Toxicity ,Agarose gel electrophoresis ,Fetal bovine serum - Abstract
COLO 201, human colon adenocarcinoma cells were incubated with artificial primers, p-nitrophenyl-glycoside derivatives at 1.0 mmol (mM) in the medium containing 10% fetal bovine serum to detect sugar chain elongation. However, when p-nitrophenyl-beta-N-acetylglucosamine (beta-GlcNAc-PNP) was added, the medium changed color to yellow and the cells were dead. To explain this finding, the cells were incubated with 1.0 mM each of beta-GlcNAc-PNP and 4-methylumbelliferyl-beta-N-acetylglucosamine, then the number of living cells was measured in a time course. In beta-GlcNAc-PNP, the living cells were decreased at 24 hours. The cells were survived with N-acetylglucosamine, whereas in the presence of p-nitrophenol (PNP) the living cells were decreased. It was suggested that PNP released from beta-GlcNAc-PNP induced the cell death. Activity of beta-D-N-acetylglucosaminidase was detected in fetal bovine serum. It was shown that PNP induced the cell death in time-and-dose dependent manner. Genomic DNA from COLO 201 analyzed by agarose gel electrophoresis was fragmentated. PNP analogues were tested for toxicity, and the results suggested that the phenolic OH-group linked to benzene ring and nitro-group linked to the structure in para-form (PNP) was the most effective.
- Published
- 2001
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