From ^3H to H3: Slide-Based Analyses of the Cell Cycle





The techniques of immunocytochemistry and <i>in situ</i> hybridization can supplant the flawed ‘mitotic index’ in providing critical information regarding cell cycle status of human tumors. While methods such as bromodeoxyuridine (BrdU) incorporation can provide accurate measurements of S phase indices in tissue samples, this method cannot generally be used for routine pathology specimens. Monoclonal and polyclonal antibodies have been developed which can, under appropriate fixation conditions, detect cells in specific phases of the cell cycle, including: the Ki-67-defined nuclear cell proliferation related antigen, expression of which is characteristic of all cycling (non-G<sub>0</sub>) cells; proliferating cell nuclear antigen (PCNA; a marker of S-phase); and statin, a marker of G<sub>0</sub>-phase. Antibodies to PCNA however, cannot be employed with reliability in deparaffinized, formalin-fixed tissues owing to the variable retention of non-repliconassociated PCNA in the nucleus in non-S-phase cells; ‘PCNA indices’, however, in methacarn fixed tissue can provide information comparable to that of BrdU incorporation. Statin is a 57kd protein, expression of which is associated with cell senescence and cell quiescence; immunolocalization is possible only in frozen or methacarn-fixed, deparaffinized tissues. Antibodies to PCNA and statin have been applied to a series of 51 methacarn breast specimens, confirming an inverse correlation between statin and PCNA indices; furthermore, a direct relationship between statin index and long term survival was observed. The most recently described marker associated with cell proliferation, <i>in situ</i> hybridization using cDNA probes to histone H3 and/or H4 mRNA, offers the unique feature of identifying a cytoplasmic S-phase marker, permitting double labeling studies with antibodies to nuclear proteins. These methods, in aggregate, represent reasonable alternatives to flow cytometrically determined cell proliferation indices.


  • Acta histochemica et cytochemica

    Acta histochemica et cytochemica 28(1), 55-59, 1995-02-01


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