Quantitation of the nucleophosmin/B23-translocation using imaging analysis.
We have previously detected by immunofluorescent assay that the cellular localization of nucleophosmin/B23 (NPM) shifts from the nucleolus to the nucleoplasm (NPM-translocation) after exposure of cells to multiple agents. In order to improve the quantification of the NPM-translocation, we have developed a digital imaging technique. Human Lo leukemia cells, MCF-7 breast carcinoma cells, and fresh human leukemia cells were exposed to anthracyclines or actinomycin D for 4 h. The degree of NPM-translocation was determined and presented as the localization index (LI). Control cells had a LI of about 10, which indicates that the majority of NPM was localized in nucleoli. The LI for drug-treated cells decreased in a dosage- and time-dependent manner. The effect of two classes of anthracycline (daunomycin and aclacinomycin A) and different types of intercalators (daunomycin and actinomycin D) had additive effects on induction of NPM-translocation. The imaging procedure was easily applied to fresh leukemia cells, thus providing useful information regarding drug effects on cancer cells.
Chan
P K
PK
Department of Pharmacology, Baylor College of Medicine, Houston, TX 77030, USA.
Qi
Y
Y
Amley
J
J
Koller
C A
CA
D015250:Aclarubicin / Q000008:administration & dosage; D000903:Antibiotics, Antineoplastic / Q000008:administration & dosage / Q000494:pharmacology; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology; D001943:Breast Neoplasms / Q000188:drug therapy / Q000378:metabolism; D002466:Cell Nucleolus / Q000378:metabolism; D003609:Dactinomycin / Q000008:administration & dosage; D003630:Daunorubicin / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005455:Fluorescent Antibody Technique; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D007948:Leukemia, Monocytic, Acute / Q000188:drug therapy / Q000378:metabolism; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism*; D000090243:Nucleophosmin; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000903:Antibiotics, Antineoplastic; C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D000090243:Nucleophosmin; D003609:Dactinomycin; D015250:Aclarubicin; D003630:Daunorubicin