Sodium butyrate induces apoptosis and accumulation of ubiquitinated proteins in human breast carcinoma cells.
To investigate the possible relationship between apoptosis and the ubiquitin pathway we examined the patterns of ubiquitinated proteins in the human breast carcinoma MCF-7 cell line following induction of apoptotic death by sodium butyrate. Apoptosis in these cells was associated with internucleosomal DNA fragmentation and proteolytic cleavage of poly(ADP-ribose) polymerase. By dual in situ antiubiquitin immunofluorescence and chromatin DNA staining, we demonstrated that ubiquitin fluorescence was increased specifically in cells that underwent sodium butyrate-mediated apoptosis. The extent of ubiquitin incorporation into protein conjugates was examined in both adherent (not yet apoptotic) and floating (apoptotic) cell populations. We found that apoptotic cells exhibited enhanced intensity of ubiquitin-immunoreactive conjugates, whereas adherent cells did not. In addition, two-dimensional immunoblot analysis of proteins from apoptotic cells identified a set of isomeric ubiquitinated conjugates located at a pI range of 4. 2 - 4.6 and a Mr approximately of 30 kDa. These data indicate that the ubiquitin pathway may play a role in the sodium butyrate-induced apoptotic program in breast carcinoma cells.
Soldatenkov
V A
VA
Department of Radiation Medicine, Division of Radiation Research, Georgetown University Medical Center, Washington DC 20007, USA. soldates@gunet.georgetown.edu
Prasad
S
S
Voloshin
Y
Y
Dritschilo
A
A
D017209:Apoptosis / Q000187:drug effects*; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology*; D020148:Butyric Acid / Q000494:pharmacology*; D053938:DNA Fragmentation / Q000187:drug effects; D005260:Female; D006801:Humans; D015151:Immunoblotting; D009363:Neoplasm Proteins / Q000302:isolation & purification / Q000378:metabolism*; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014452:Ubiquitins / Q000378:metabolism*
D009363:Neoplasm Proteins; D014452:Ubiquitins; D020148:Butyric Acid; D011065:Poly(ADP-ribose) Polymerases