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Temporal Tracking of Cell Cycle Progression Using Flow Cytometry without the Need for Synchronization
Published on: August 16, 2015
Cell cycle kinetics in malignant lymphoma studied with in vivo iododeoxyuridine administration, nuclear Ki-67
P P Brons1, J M Raemaekers, M J Bogman
1Department of Hematology, University Hospital Nijmegen, The Netherlands.
Insights
This study reveals that in vivo labeling with iododeoxyuridine (IdUrd) and flow cytometry (FCM) can detail malignant lymphoma cell cycle kinetics. Cell cycle phase durations vary significantly, even within similar lymphoma subtypes.
Area of Science:
- Oncology
- Cell Biology
- Biophysics
Background:
- Understanding cell cycle kinetics is crucial for diagnosing and treating malignant lymphomas.
- Traditional methods for assessing cell proliferation in lymphomas are often complex and require multiple biopsies.
Purpose of the Study:
- To investigate the cell cycle kinetics of malignant lymphoma using a novel in vivo labeling and flow cytometry approach.
- To establish a comprehensive cell kinetic profile from a single tumor biopsy.
Main Methods:
- In vivo labeling of lymphoma cells with iododeoxyuridine (IdUrd).
- Subsequent analysis using flow cytometry (FCM) to measure IdUrd/DNA and Ki-67/DNA.
- Studied tissue samples from 33 patients with non-Hodgkin's lymphoma (NHL), Hodgkin's disease (HD), and reactive hyperplasia.
Main Results:
- The approach provided extensive cell kinetic data including S-phase percentage, IdUrd labeling index (LI), growth fraction, and cell cycle phase durations.
- In NHL, S-phase percentage, LI, growth fraction, S-phase duration, and cell production rate correlated significantly with histologic malignancy grade.
- Cell cycle phase durations (G1, S) and total cell cycle time were largely independent of malignancy grade within NHL.
- IdUrd LI strongly correlated with S-phase percentage, growth fraction, potential doubling time, and cell production rate.
Conclusions:
- Detailed in vivo growth characteristics of malignant lymphoma can be obtained using IdUrd labeling and FCM.
- The transition time through different cell cycle phases exhibits significant variability, even within specific lymphoma histologic subgroups.
Abstract:
Cell cycle kinetics of malignant lymphoma were investigated using in vivo labeling with iododeoxyuridine (IdUrd) and subsequent flow cytometry (FCM) of IdUrd/DNA and Ki-67/DNA. This approach provides an extensive cell kinetic profile from only one single tumor biopsy, including data upon the percentage of S-phase cells, the IdUrd labeling index (LI), Ki-67-derived growth fraction, duration of the S-phase, duration of the G1-phase, potential doubling time, cell production rate, and total cell cycle time. Tissue samples from 33 patients were studied: non-Hodgkin's lymphoma (NHL; n = 22), Hodgkin's disease (HD; n = 7), and reactive hyperplasia (n = 4). In NHL, the percentage of S-phase cells, LI, growth fraction, duration of the S-phase, and cell production rate were significantly correlated with the histologic malignancy grade according to the Working Formulation (P < or = .02). Data found in HD were not essentially different from those in low-grade NHL and reactive hyperplasia. Remarkably, the duration of the S-phase, the duration of the G1-phase, and the total cell cycle time appeared to be rather independent of histologic malignancy grade within the NHL category. A significant correlation was observed between the IdUrd LI and the percentage of S-phase cells, the growth fraction, the potential doubling time, and the cell production rate (P < .001), but not with the duration of the separate cell cycle phases (P > .05). Our data show (1) that it is feasible to obtain detailed information on the in vivo growth characteristics of malignant lymphoma; and (2) that the transition time through the different cell cycle phases widely varies, even within distinct histologic subgroups.
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