Premium
Fludarabine nucleoside induces accumulations of p53, p63 and p73 in the nuclei of human B ‐lymphoid cell lines, with cytosolic and mitochondrial increases in p53
Author(s) -
Almazi Juhura G.,
Mactier Swetlana,
Best O. Giles,
Crossett Ben,
Mulligan Stephen P.,
Christopherson Richard I.
Publication year - 2012
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201200003
Subject(s) - raji cell , microbiology and biotechnology , gene isoform , fludarabine , apoptosis , biology , blot , cytosol , cell culture , mitochondrion , chemistry , cell , biochemistry , gene , genetics , enzyme , chemotherapy , cyclophosphamide
Purpose Human R aji cells treated with fludarabine nucleoside (2‐ F ara A , 3 μM) undergo apoptosis with accumulation of p53 in the nuclei as multiple phosphorylated isoforms and C ‐terminal truncated derivatives. Changes induced by 2‐ F ara A in the levels of p53, p63 and p73 in the nuclear, cytosolic and mitochondrial fractions have been determined in four human B ‐lymphoid cell lines that are TP 53‐functional ( R aji and IM 9) and TP 53‐mutated ( MEC 1 and U 266). Experimental design The B ‐lymphoid cell lines were treated with 2‐ F ara A (3 μM, 24 h, 48 h) and viability determined. Protein extracts of subcellular fractions from 2‐ F ara A ‐treated cells were analysed by 1D and 2D electrophoresis; multiple phosphorylated isoforms and truncated derivatives were identified by W estern blots for p53, p63 and p73. Results p53 and p63 were present in all three fractions, while p73 was only detected in nuclei. After treatment with 2‐ F ara A , nuclear p53, p63 and p73 accumulated as multiple phosphorylated isoforms and truncated derivatives. The association of p63 with mitochondria in human cells is novel. Conclusions and clinical relevance Comprehensive information on the subcellular distributions and responses of p53, p63 and p73 to 2‐ F ara A provides additional insight into mechanisms for induction of apoptosis in the treatment of B ‐lymphoproliferative disorders with fludarabine.