Evaluation of the relative contribution of p53-mediated pathway in X-ray-induced apoptosis in human leukemic MOLT-4 cells by transfection with a mutant p53 gene …

H Nakano, M Kohara, K Shinohara - Cell and tissue research, 2001 - Springer
H Nakano, M Kohara, K Shinohara
Cell and tissue research, 2001Springer
There are several pathways leading to apoptosis. It is not clear whether cells choose one of
them or use multiple processes when they commit to apoptosis. MOLT-4 cells undergo
apoptosis after X-irradiation through the p53-dependent pathway and/or ceramide signal. To
evaluate the relative contribution of these pathways, we studied effects of the expression of
various levels of transfected murine mutant p53 cDNA (TGC→ CGC of codon 173,
corresponding to codon176 in human p53) on the induction of apoptosis in X-irradiated or …
Abstract
There are several pathways leading to apoptosis. It is not clear whether cells choose one of them or use multiple processes when they commit to apoptosis. MOLT-4 cells undergo apoptosis after X-irradiation through the p53-dependent pathway and/or ceramide signal. To evaluate the relative contribution of these pathways, we studied effects of the expression of various levels of transfected murine mutant p53 cDNA (TGC→CGC of codon 173, corresponding to codon176 in human p53) on the induction of apoptosis in X-irradiated or heated MOLT-4 cells. When survival was determined by the dye-exclusion test at 24 h after irradiation, the percentage of X-ray- or heat-induced dead cells was markedly decreased, depending on the expression level of mutant p53 protein in transfected clones. The appearance of apoptotic cells as determined by morphological changes was also decreased. These inhibitions were almost complete at 24 h after irradiation with X-rays in the case of the highest-expressing clone. p21WAF1 protein was increased in MOLT-4 cells after X-irradiation, but not in the transfectant. These results suggest that murine mutant p53 protein has a dominant-negative effect against normal p53 in MOLT-4, and that the X-ray-induced apoptosis in MOLT-4 is fully p53-dependent.
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