Comparison of bromodeoxyuridine labeling indices obtained from tissue sections and flow cytometry of brain tumors

Abstract

Sixteen patients with brain tumors were given a 30- to 60-minute intravenous infusion of bromodeoxyuridine (BUdR), 200 mg/sq m. Grossly viable fragments were taken from the biopsied tumor specimens and divided into two portions. One portion was dissociated into single cells, stained both with fluorescein isothiocyanate (FITC) using anti-BUdR monoclonal antibody as the first antibody and with propidium iodide (for deoxyribonucleic acid), and analyzed by flow cytometry (FCM). The labeling index (LI) was calculated as the number of FITC-labeled cells expressed as a percentage of the total number of cells analyzed. The other portion was fixed in 70% ethanol, embedded in paraffin, sectioned, and stained with immunoperoxidase using anti-BUdR monoclonal antibody as the first antibody. The LI of these tissue sections was calculated in two ways: from selected areas in which the labeled cells were evenly distributed and from the entire tissue section. The LI's obtained by FCM correlated closely with those from the entire tissue sections (r = 0.99, p less than 0.000001) and were usually lower than LI's from selected areas of tissue sections. The LI's determined by FCM also correlated well with the LI's from selected areas of tissue sections (r = 0.82, p less than 0.00012), despite the difference in values between them. Thus, the FCM-derived LI and the tissue LI can both provide useful information for predicting the biological malignancy of individual tumors and for designing treatment regimens for individual patients with brain tumors; however, different standards should be used to interpret the LI's obtained by these two methods.

Modulation of BUdR labeling index in rat brain tumors following intracarotid ACNU administration

“…We therefore evaluate the changes of proliferating activity of intracerebrally transplanted brain tumors using BUdR and anti-BUdR monoclonal antibody [9,10]. Intracarotid ACNU decreased the LI markedly in sensitive tumors and sustained the low LI for a relatively long time.…”

Cell-Kinetic Characteristics of Human Brain Tumors

“…Since medullo blastomas are the tumors with the highest LI [30] we observed that the tumor studied was a 5-year recur rence of a medulloblastoma first operated in a 30-yearold patient and treated with radio-and chemotherapy. These atypical clinical features and the adjuvant treat ment might explain the kinetic results; infact, it has been previously reported that medulloblastomas of adult age show a less agressive growth and that the adjuvant treatment can select specific clones with individual biological characteristics [40].…”

“…Recently. Nagashima et al [30] studied the reliability of the BUDR-LI obtained by slides exarni-nation and by biparametric FCM showing that the BUDR-LI obtained on selected portions of slide is a more reliable method of calculation of the prolifera tive characteristics of brain tumors, assuming that the BUDR-LI determined by FCM is a very good method and if a different magnitude of values is employed for prognostic purposes. Since the biparametric FCM is a faster method, it may be routinely used for the deter mination of the BUDR-LI of malignant brain tumors.…”

“…The evaluation of cell cycle parameters by flow cytometric measurement of bromodeoxyuridine (BUDR) incorporation into DNA has been described as a useful tool for the safe and fast determination of growth characteristics of human brain tumors [8,22,30], We have previously shown that it is possible to measure cell kinetics of human brain tumors with flow cytometry (FCM) after an in vivo injection of BUDR. The main advantages of this flow cytometric tech nique are that it is really fast (the results can be obtained within a day of surgery), it can be used in vivo (since BUDR is not radioactive or toxic at the doses required for cell-kinetic studies), it makes pos sible the estimation of both the labelling index (LI) and the duration of S-phase (TS).…”

Prognostic Value of S-Phase Fraction and DNA Ploidy in Oral Carcinoma Studied with Bromodeoxyuridine Labeling in vitro and Flow Cytometry

“…Hence, this technique permits a selective assessment of proliferation of tumor cell populations alone, provided they arc aneuploid. The utility of this procedure has been demonstrated in patients with various types of solid tumors who received BrdUrd intra venously [17,18,25]. For clinical routine application, of course, BrdUrd labeling in vitro should be preferred [5], This study is the first demonstrat ing bivariate BrdUrd/DNA flow cytometry in a homogeneous group of patients with primary and prviously untreated squa mous cell carcinomas of the oral cavity.…”