Quantitation of the nucleophosmin/B23-translocation using imaging analysis

Cancer Lett. 1996 Feb 27;100(1-2):191-7. doi: 10.1016/0304-3835(95)04100-1.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Aclarubicin / administration & dosage
  • Antibiotics, Antineoplastic / administration & dosage
  • Antibiotics, Antineoplastic / pharmacology
  • Antineoplastic Combined Chemotherapy Protocols / pharmacology
  • Breast Neoplasms / drug therapy
  • Breast Neoplasms / metabolism
  • Cell Nucleolus / metabolism
  • Dactinomycin / administration & dosage
  • Daunorubicin / pharmacology
  • Dose-Response Relationship, Drug
  • Fluorescent Antibody Technique
  • Humans
  • Image Processing, Computer-Assisted / methods*
  • Leukemia, Monocytic, Acute / drug therapy
  • Leukemia, Monocytic, Acute / metabolism
  • Nuclear Proteins / analysis
  • Nuclear Proteins / metabolism*
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / drug therapy
  • Precursor Cell Lymphoblastic Leukemia-Lymphoma / metabolism
  • Tumor Cells, Cultured

Substances

  • Antibiotics, Antineoplastic
  • Nuclear Proteins
  • nucleophosmin
  • Dactinomycin
  • Aclarubicin
  • Daunorubicin