Cell cycle-related proteins: a flow cytofluorometric study in human tumors

Marco Danova, Alberto Riccardi, Monica Giordano, Margherita Girino, Giuliano Mazzini, Laura Dezza, Edoardo Ascari

Research output: Contribution to journalArticlepeer-review

Abstract

We used 2-parameter flow cytometry (FCM) to investigate the relationship between the cell cycle phases and 3 proteins whose expression is known to increase in proliferating cells: the surface antigen transferrin receptor Trf-r), the "cyclin" (a proliferating cell nuclear antigen, PCNA), and the nuclear antigen recognized by the monoclonal antibody (MoAb) Ki-67. FITC-labeled antibodies against Trf-r, PCNA, and the Ki-67-reactive antigen, as well as propidium iodide-DNA distribution, were simultaneously measured on human leukemia HL-60 and K562, and breast carcinoma MCF-7 cell lines and on fresh human leukemic and glioblastoma cells. The 70% ethanol fixation for Trf-r and PCNA and the 95% acetone fixation for Ki-67 plus permeabilization (with 0.1% and 1% Triton X100, respectively, for the surface and the nuclear antigens) produced cell suspensions with negligible cell clumping, high-quality DNA profiles, and bright specific immunofluorescent staining. The investigated proteins have different relationships with the proliferative state of the cell. Trf-r is expressed mainly at the transition from G0/G1 to S-phase. PCNA expression is prominent in late G1 and through S-phase and decreases in G2-M. The Ki-67-reactive antigen is widely distributed in G1, S, and G2-M phases. Knowledge regarding the relationships between proliferation-associated antigens and cell cycle phase in normal and neoplastic cells could improve our understanding of the mechanisms underlying growth regulation and neoplastic transformation. Bivariate FCM is an easy method for obtaining these data from large numbers of cells.

Original languageEnglish
Pages (from-to)23-28
Number of pages6
JournalBiology of the cell / under the auspices of the European Cell Biology Organization
Volume64
Issue number1
Publication statusPublished - 1988

Keywords

  • cell proliferation
  • flow cytometry
  • monoclonal antibodies
  • surface and nuclear proteins

ASJC Scopus subject areas

  • Cell Biology

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