Cell kinetic studies of in situ human brain tumors with bromodeoxyuridine.
At the time of surgery, 18 patients with various brain tumors were given a 1-h i.v. infusion of bromodeoxyuridine (BrdUrd), 150-200 mg/m2. At an infusion rate of 200 mg/m2/h, serum BrdUrd levels of 8 microM were achieved. After the infusion, tumor tissue was obtained and divided into two portions. One portion was fixed in 70% ethanol, embedded in paraffin, and sectioned; the sections were deparaffinized, denatured with 2 N HCl, and reacted with monoclonal antibodies against BrdUrd (anti-BrdUrd MAb). BrdUrd-labeled nuclei were demonstrated satisfactorily by an indirect peroxidase method. The other portion was dissociated into single cells with a DNase enzyme cocktail and reacted with FITC-conjugated anti-BrdUrd MAb to determine the percentage of BrdUrd-labeled cells or with chromomycin A3 for DNA analysis. The single-cell suspensions were analyzed by flow cytometry. The fraction of S-phase cells in the tissue sections was similar to both the percentage of BrdUrd-labeled nuclei and the S-phase fraction determined by flow cytometric analysis. The results obtained with BrdUrd-labeled nuclei were similar to those obtained from previous autoradiographic studies of various brain tumors exposed to a pulse of 3H-thymidine. Since BrdUrd is not radioactive and is nontoxic at the dosage used, these techniques, together with the histopathological diagnosis, may help to predict the biological malignancy of individual tumors.
D000911:Antibodies, Monoclonal; D001254:Astrocytoma / Q000473:pathology*; D001932:Brain Neoplasms / Q000473:pathology*; D001973:Bromodeoxyuridine / Q000097:blood; D002453:Cell Cycle; D004273:DNA, Neoplasm / Q000032:analysis* / Q000096:biosynthesis; D005434:Flow Cytometry; D005909:Glioblastoma / Q000473:pathology*; D006801:Humans; D007124:Immunoenzyme Techniques; D007399:Interphase; D008579:Meningioma / Q000473:pathology*; D008657:Metabolic Clearance Rate
D000911:Antibodies, Monoclonal; D004273:DNA, Neoplasm; D001973:Bromodeoxyuridine