Cell kinetic studies of in situ human brain tumors with bromodeoxyuridine

Hoshino, Takao; Nagashima, Tadashi; Murovic, Judith A.; Levin, Ellen M.; Levin, Victor A.; Rupp, Stephen M.

doi:10.1002/cyto.990060619

Cited by 151 publications

(40 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This occurred in 6/17 non-malignant and in 6/33 malignant specimens, indicating that successful analysis is possible in an even higher percentage of patients with malignant tumours. Acute side-effects were not observed with the dose of BrdU employed, as has also been the case in earlier studies with this dose and slightly higher doses (Forster et al, 1992;Hoshino et al, 1985;Hupperets et al, 1986;Miller et al, 1991;Nagashima et al, 1988;Shimomatsuya et al, 1991;Ten Velde et al, 1989a;Wilson et al, 1988).…”

Section: Discussionsupporting

confidence: 74%

Cytokinetic analysis of lung cancer by in vivo bromodeoxyuridine labelling

1994

Lung Cancer

View full text Add to dashboard Cite

Summary Cytokinetic parameters of various types of lung cancer were determined in bronchoscopy specimens after in vivo labelling with the thymidine analogue bromodeoxyuridine (BrdU). The S-phase fraction and BrdU labelling index were measured flow cytometrically, allowing calculation of the S-phase transit time and potential tumour doubling time. The methodology used was found to be feasible for obtaining cytokinetic data from 76% of the bronchial biopsy samples.Despite the difference in clinical behaviour and growth pattern between small cell lung cancer (SCLC) and non-small cell lung cancer (NSCLC), no significant differences were observed between the mean values of the cytokinetic parameters of SCLC and NSCLC. The estimated cell loss factor was higher in NSCLC than in SCLC.It appears that the growth of a tumour, as clinically observed, is to a considerable extent influenced by cell loss. In accord with this assumption is the fact that we have observed non-BrdU labelled S-phase cells, both in tumour biopsies and in apparently normal tissue. The presence of these so-called unlabelled S-phase cells in relation to cell loss is discussed.Individual lung cancers differ in histological pattern and other phenotypic characteristics. These differences are relevant in predicting therapeutic responsiveness and prognosis (Fraire et al.,1992). In recent years it has become evidence that in lung cancer as well as in other types of malignancy, cell cycle parameters may be important prognosticators (Tubiana & Courdi, 1989). In several studies, a high S-phase fraction (SPF) was found to be associated with a shorter survival time (Ten Velde et al., 1988;Volm et al., 1985); this was also found for tumours other than lung carcinoma, including breast carcinoma (Kallioniemi et al., 1986;O'Reilly et al., 1990). In addition, a low ex vivo thymidine labelling index in patients with stage I NSCLC predicts a longer survival time (Silvestrini et al., 1991 informed consent, the patients were infused with 50 mg m-2 BrdU (Janssen Pharmaceutica, Beerse, Belgium), dissolved in 100 ml 0.9% NaCI, within a timespan of 10 min. The BrdU was given approximately 4 to 5 h before bronchoscopy. Approval for the in vivo labelling method was given by the ethical committee of the University Hospital of Maastricht. Biopsies were taken with a flexible bronchoscope, fixed in formalin for routine diagnosis and in 70% ethanol for flow cytometric analysis. The latter samples were stored at 4°C until use. Flow cytometry and BrdU detectionThe biopsy specimens were double-stained with anti-BrdU (clone IIbS; Schutte et al., 1987) and propidium iodide (PI), using the protocol described by Schutte et al. (1987). Briefly, ethanol fixed biopsies were minced in a petri dish and washed twice in phosphate buffered saline (PBS) pH 7.4, by centrifugation for 5 min at 400 g. To obtain nuclei, the cell suspension was digested with 0.4 mg ml-' pepsin (Boehringer Mannheim, Germany; 108057) in 0.1 N HCI for 30 min at room temperature. Undigested fragments were then removed by si...

show abstract

Section: Discussionsupporting

confidence: 74%

Cytokinetic analysis of lung cancer by in vivo bromodeoxyuridine labelling

1994

Lung Cancer

View full text Add to dashboard Cite

show abstract

“…Our BrdUrdiDNA procedure for detecting resistance to Ara-C is attractive for several reasons: 1) It is rapid, giving results within a few hours of sample acquisition; 2) the assay can be directed toward cells with S-phase DNA content, so variable S-phase fractions in tumor populations can be accounted for; 3) the procedure can be directed toward tumor cells in a mixed population as markers for these tumor cells become available; and 4) it can determine the frequency of rare resistant cells quantitatively. In addition, BrdUrd is sufficiently nontoxic that it may be administered to leukemic patients (15,9).…”

Section: Discussionmentioning

confidence: 99%

Detection of cytosine arabinoside resistant cells at low frequency using the bromodeoxyuridine/DNA assay

1985

View full text Add to dashboard Cite

This report describes a rapid and sensitive procedure for detection of cytosine arabinoside‐ (Ara‐C) resistant mouse leukemia cells (L1210) in a predominantly Ara‐C‐sensitive population. L1210 cell lines sensitive or resistant to Ara‐C were grown and treated with Ara‐C in vitro or in vivo. Ara‐C‐resistant cells were detected as those cells with S‐phase DNA content retaining the ability to incorporate bromodeoxyuridine (BrdUrd) after treatment with Ara‐C. The BrdUrd incorporation ability of the S‐phase cells was assessed by simultaneous flow cytometric measurement of cellular DNA content and amount of incorporated BrdUrd. The proportion of Ara‐C‐resistant cells was accurately estimated at frequencies approaching 10−3.

show abstract

“…Measurements of incorporation of bromodeoxyuridine (BrdUrd) into DNA have provided a useful assessment of DNA synthesis in leukemia cells in vitro and in vivo (3)(4)(5)8,9,12). The amount of BrdUrd incorporated into DNA can be quantified by either flow cytometric (3)(4)(5)12) or microscopic techniques (8,9).…”

mentioning

confidence: 99%

“…The amount of BrdUrd incorporated into DNA can be quantified by either flow cytometric (3)(4)(5)12) or microscopic techniques (8,9). If BrdUrd incorporation is to be a reliable in vivo measure, possible concentration-dependent artifacts must be avoided.…”

mentioning

confidence: 99%

“…It is expected that similar variations in plasma concentrations of other nucleosides such as BrdUrd might occur. BrdUrd doses (200 mgim' over 1 h) used in in vivo cytokinetic studies yield concentrations in the 8 pM range (5). A recent trial employed a lower dose of 100 mgim' (8); although pharmacokinetics were not evaluated in this study, elimination is expected to be linear (10).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Bromodeoxyuridine incorporation into DNA of human leukemia cells is not concentration dependent

et al. 1990

View full text Add to dashboard Cite

Leukemia blasts isolated from bone marrow aspirates of 44 adults with acute leukemia were incubated for 1 h with 0.008-32 pM bromodeoxyuridine (BrdUrd). After dual labeling with monoclonal anti-BrdUrd antibodies and propidium iodide, the cells were analyzed by flow cytometry. Percent labeled cells and intensity of labeling were similar over concentrations of BrdUrd ranging from 0.8-32 pM-a 40-fold range. Therefore, despite potential interpatient variability in nucleoside pharmacokinetics, commonly used doses of BrdUrd which are intended to achieve steady-state plasma concentrations in the 8.0 pM range can be expected to provide a reliable estimate of the S-phase fraction. Key terms: Cell kinetics, acute leukemia, concentration independence, flow cytometryMeasurements of incorporation of bromodeoxyuridine (BrdUrd) into DNA have provided a useful assessment of DNA synthesis in leukemia cells in vitro and in vivo (3)(4)(5)8,9,12). The amount of BrdUrd incorporated into DNA can be quantified by either flow cytometric (3-5,12) or microscopic techniques (8,9). If BrdUrd incorporation is to be a reliable in vivo measure, possible concentration-dependent artifacts must be avoided. Although interpatient variations in BrdUrd pharmacokinetics have not been extensively evaluated, continuous infusions of a fixed dose of the nucleoside cytosine arabinoside in leukemia patients can result in a 10-fold variation in plasma concentration a t steady state (1). It is expected that similar variations in plasma concentrations of other nucleosides such as BrdUrd might occur. BrdUrd doses (200 mgim' over 1 h) used in in vivo cytokinetic studies yield concentrations in the 8 pM range (5). A recent trial employed a lower dose of 100 mgim' (8); although pharmacokinetics were not evaluated in this study, elimination is expected to be linear (10). The toxicities of BrdUrd are dose dependent, but are not expected with the doses administered for cytokinetic studies (6).In an attempt to establish a range of concentrations over which BrdUrd incorporation into DNA of human leukemia cells is constant, the following experiment was performed. MATERIALS AND METHODSIsolation of Leukemia Cells A separate marrow aspirate from the posterior iliac crest of adult patients with acute leukemia was collected in heparinized syringes and resuspended in 30 cc of ice-cold RPMI medium 1640 (Gibco, Grand Island, NY). This suspension was layered over 15 ml of FicollPaque (Pharmacia, Picataway, NJ; density = 1.077) and centrifuged at 1,OOOg for 20 min. Cells at the plasma-Ficoll interface were harvested and washed twice with 50 ml of RPMI and then resuspended in 2 ml of RPMI with 10% fetal calf serum (FCS)

show abstract

Cell kinetic studies of in situ human brain tumors with bromodeoxyuridine

Cited by 151 publications

References 18 publications

Cytokinetic analysis of lung cancer by in vivo bromodeoxyuridine labelling

Cytokinetic analysis of lung cancer by in vivo bromodeoxyuridine labelling

Detection of cytosine arabinoside resistant cells at low frequency using the bromodeoxyuridine/DNA assay

Bromodeoxyuridine incorporation into DNA of human leukemia cells is not concentration dependent

Contact Info

Product

Resources

About