Nucleolar protein B23 translocation after deferoxamine treatment in a human leukemia cell line.
Localization of nucleolar protein B23 in HL-60 cells under the treatment by iron chelator deferoxamine (DSF) was studied using indirect immunofluorescence. Bright nucleolar fluorescence was observed in exponentially growing control cells. The addition of DSF in the culture system resulted in time- and dose-dependent induction of protein B23 translocation from nucleoli to nucleoplasm, inhibition of cell growth, DNA and RNA synthesis. The addition of FeCl3 at culture initiation completely reversed the effects of DSF. Furthermore, significant numbers of HL-60 cells could be rescued from the effects of DSF when iron was added back as late as 24 hr after exposure to DSF. Cells resumed their abilities to grow and to synthesize DNA and RNA upon the iron rescue. Protein B23, accordingly, relocated from nucleoplasm to nucleoli. These results indicate the importance of iron for proliferation of leukemic cells and localization of protein B23 in nucleoli. Preribosomal ribonucleoprotein particles (pre-rRNPs) were extracted from isolated nucleoli of HL-60 cells and fractionated on sucrose density gradients. Protein B23 was found to be co-localized with the pre-rRNPs as determined by ELISA assays. No such B23-associated pre-rRNPs or other pre-rRNP fractions were obtained from nucleoli of DSF-treated cells. These results suggest that one of the effects of the anti-proliferative action of DSF is the inhibition of rRNA synthesis in nucleoli. Due to the lack of new synthesis of rRNA in nucleoli, protein B23 loses its binding target and translocates into the nucleoplasm. B23 translocation, as observed by immunofluorescence, may be a simple and rapid method for assessing inhibition of cell growth in response to anti-proliferative drugs such as deferoxamine in cancer chemotherapy.
Yung
B Y
BY
Department of Pharmacology, Chang Gung Medical College, Taiwan, Republic of China.
Yang
Y H
YH
Bor
A M
AM
D000911:Antibodies, Monoclonal; D002455:Cell Division; D002460:Cell Line; D002466:Cell Nucleolus / Q000378:metabolism* / Q000648:ultrastructure; D002467:Cell Nucleus / Q000378:metabolism*; D002712:Chlorides; D004261:DNA Replication / Q000187:drug effects; D003676:Deferoxamine / Q000494:pharmacology*; D005290:Ferric Compounds / Q000494:pharmacology; D005455:Fluorescent Antibody Technique; D006801:Humans; D015473:Leukemia, Promyelocytic, Acute; D009687:Nuclear Proteins / Q000378:metabolism*; D000090243:Nucleophosmin; D010750:Phosphoproteins / Q000378:metabolism; D012322:RNA Precursors / Q000235:genetics / Q000378:metabolism; D012334:RNA, Neoplasm / Q000096:biosynthesis / Q000187:drug effects; D012335:RNA, Ribosomal / Q000096:biosynthesis; D014529:Uridine / Q000378:metabolism
D000911:Antibodies, Monoclonal; D002712:Chlorides; D005290:Ferric Compounds; D009687:Nuclear Proteins; D010750:Phosphoproteins; D012322:RNA Precursors; D012334:RNA, Neoplasm; D012335:RNA, Ribosomal; D000090243:Nucleophosmin; D003676:Deferoxamine; C024555:ferric chloride; D014529:Uridine