The mechanism of cell death in human cultured colon adenocarcinoma cell line COLO 201 induced by beta-D-N-acetylglucosaminyl-p-nitrophenol.
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.
Kukidome
J
J
Department of Biochemistry, Hirosaki University School of Medicine, Japan.
Kakizaki
I
I
Takagaki
K
K
Matsuki
A
A
Munakata
A
A
Endo
M
M
D000117:Acetylglucosamine / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology*; D000230:Adenocarcinoma; D017209:Apoptosis; D018929:Cell Culture Techniques; D003110:Colonic Neoplasms; D004305:Dose-Response Relationship, Drug; D006801:Humans; D015394:Molecular Structure; D009596:Nitrophenols / Q000737:chemistry / Q000494:pharmacology*; D013997:Time Factors; D014407:Tumor Cells, Cultured
D009596:Nitrophenols; C014124:4-nitrophenyl-N-acetyl-2-deoxyglucopyranoside; D000117:Acetylglucosamine