Detection of proliferating S-phase brain tumor cells by in situ DNA replication.
OBJECTIVE
Current methods used to describe the proliferative status of brain tumors rely on labor-intensive, potentially costly procedures. This article provides a description of a rapid, inexpensive, uncomplicated technique used to identify proliferating cells in tissue obtained at the time of resection.


METHODS
Touch preparations of 16 fresh astrocytic tumors and four fresh healthy temporal neocortical tissue samples were obtained at the time of surgery. Slides were placed in hypotonic potassium chloride to permeabilize their membranes, incubated in nucleotide precursors, and labeled with bromodeoxyuridine; they were later examined with the aid of a fluorescence microscope. The percentage of tumor cells in the S phase increased in conjunction with the grade of tumor and corresponded with the findings of immunohistochemical staining for the cell-cycle marker MIB-1. These results were confirmed in cell culture by using normal human astrocytes and two glioma cell lines. Slides can be analyzed in as little as 30 minutes after removal of tissue during surgery.


CONCLUSIONS
In this study the authors describe a simple method by which cells in the S phase of the cell cycle. which are contained in fresh tumor obtained at the time of surgery, can be labeled. This method may prove a useful adjunct to frozen-section analysis and may permit discrimination of neoplastic tissues from other tissues observed in small specimen samples.
Weil
R J
RJ
Department of Neurosurgery, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2380, USA. robert.weil@surgery.mc.vanderbilt.edu
Toms
S A
SA
Johnson
M D
MD
Mealer
A
A
D034961:Antigens, Nuclear; D001254:Astrocytoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D001932:Brain Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology*; D002455:Cell Division; D002478:Cells, Cultured; D004261:DNA Replication; D005909:Glioblastoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D019394:Ki-67 Antigen; D019579:Neocortex / Q000166:cytology / Q000502:physiology; D009462:Neurology / Q000379:methods*; D009687:Nuclear Proteins / Q000378:metabolism; D012016:Reference Values; D016196:S Phase; D013997:Time Factors
D034961:Antigens, Nuclear; D019394:Ki-67 Antigen; D009687:Nuclear Proteins