Summary
We compared the flow cytometric measurement and analysis of the potential doubling time (T pot) between three centres involved in the National Cancer Institute (NCI) protocol T92-0045. The primary purpose was to understand and minimize the variation within the measurement. A total of 102 specimens were selected at random from patients entered into the trial. Samples were prepared, stained, run and analysed in each centre and a single set of data analysed by all three centres. Analysis of the disc data set revealed that the measurement of labelling index (LI) was robust and reproducible. The estimation of duration of S-phase (T s) was subject to errors of profile interpretation, particularly DNA ploidy status, and analysis. The LI dominated the variation in T pot such that the level of final agreement, after removal of outliers and ploidy agreement, reached correlation coefficients of 0.9. The sample data showed poor agreement within each of the components of the measurement. There was some improvement when ploidy was in agreement, but correlation coefficients failed to exceed values of 0.5 for T pot. The data suggest that observer-associated analysis of T s and tissue processing and tumour heterogeneity were the major causes of variability in the T pot measurement. The first two aspects can be standardized and minimized, but heterogeneity will remain a problem with biopsy techniques.
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References
Begg, A. C. (1995). The clinical status of Tpot as a predictor? Or why no tempest in the Tpot!. Int J Radiat Oncol Biol Phys 32: 1539–1541.
Begg, A. C., Hofland, I., Moonen, L., Bartelink, H., Schraub, S., Bontemps, P., Le Fir, R., Van den Bogaert, W., Caspers, R., van Glabbeke, M. & Horiot, J-C (1990). The predictive value of cell kinetic measurements in a European trial of accelerated fractionation in advanced head and neck tumours: an interim report. Int J Radiat Oncol Biol Phys 19: 1449–1453.
Begg, A. C., Moonen, L., Hofland, I., Dessing, M. & Bartelink, H. (1988). Human tumour cell kinetics using a monoclonal antibody against iododeoxyuridine; intratumour sampling variations. Radiother Oncol 11: 337–347.
Begg, A. C., McNally, N. J., Shrieve, D. C. & Karcher, H. A. (1985). A method to measure the DNA synthesis and the potential doubling time from a single sample. Cytometry 6: 620–626.
Bennett, M. H., Wilson, G. D., Dische, S., Saunders, M. I., Martindale, C. A. & Robinson, B. M. (1992). Tumour proliferation assessed by combined histological and flow cytometric analysis: implications for therapy in squamous cell carcinoma in the head and neck. Br J Cancer 65: 870–878.
Bland, M. J. & Altman, D. G. (1986). Statistical methods for assessing agreement between two methods of clinical measurement. Lancet i: 307–310.
Bourhis, J., Wilson, G., Wibault, P., Bosq, J., Chauvaudra, N., Janot, F., Luboinski, B., Eschwege, F. & Malaise, E. P. (1993). In vivo measurement of potential doubling time by flow cytometry in oropharyngeal cancer treated by conventional radiotherapy. Int J Radiat Oncol Biol Phys 26: 793–799.
Corvo, R., Giaretti, W., Sanguinsti, G., Geido, E., Orecchia, R., Barra, S., Margarino, G., Bacigalupo, A. & Vitale, V. (1993). Potential doubling time in head and neck tumours treated by primary radiotherapy – preliminary evidence for a prognostic significance in local control. Int J Radiat Oncol Biol Phys 27: 1165–1172.
Dische, S., Saunders, M., Barrett, A., Harvey, A., Gibson, D. & Parmar, M. (1997). A randomised multicentre trial of CHART versus conventional radiotherapy in head and neck cancer. Radiother Oncol 44: 123–136.
Fowler, J. F. (1990). How worthwhile are short schedules in radiotherapy? A series of exploratory calculations. Radiother Oncol 18: 165–181.
Fowler, J. F. & Lindstrom, M. (1992). Loss of local control with prolongation in radiotherapy. Int J Radiat Oncol Biol Phys 23: 457–467.
Haustermans, K., Hofland, I., Pottie, G., Ramaekers, M. & Begg, A. C. (1995). Can measurements of potential doubling time (Tpot) be compared between laboratories? A quality control study. Cytometry 19: 154–163.
Horiot, J. C., Bontemps, P., van den Bogaert, W., Le Fur, R., van den Weijngaert, D., Bolla, M., Bernier, J., Lusinchi, A., Stuschke, M., Lopez-Torrecilla, J., Begg, A. C., Pierart, M. & Collette, L. (1997). Accelerated fractionation (AF) compared to conventional fractionation (CF) improves loco-regional control in the radiotherapy of advanced head and neck cancers: results of the EORTC 22851 randomized trial. Radiother Oncol 44: 111–121.
Peters, L. J., Ang, K. K. & Thames, H. J. (1988). Accelerated fractionation in the radiation treatment of head and neck cancer. A critical comparison of different strategies. Acta Oncol 27: 185–194.
Shackney, S. E. & Shankey, T. V. (1995). Genetic and phenotypic heterogeneity of human malignancies: Finding order in chaos. Cytometry 21: 2–5.
Shankey, T. V., Rabinovitch, P. S., Bagwell, B., Bauer, K. B., Duque, R. E., Hedley, D. W., Mayall, B. H., Wheeless, L. & Cox, C. (1993). Guidelines for implementation of clinical DNA cytometry. Cytometry 14: 472–477.
Steel, G. G. (1977). Growth Kinetics of Tumours, Clarendon Press: Oxford
Terry, N. H. A. & Peters, L. J. (1995). The predictive value of tumour cell kinetic parameters in radiotherapy: Considerations regarding data production and analysis. J Clin Oncol 13: 1833–1836.
Tsang, R. W., Fyles, A. W., Kirkbride, P., Levin, W., Manchul, L. A., Rawlings, G. A., Banerjee, M., Pintillie, M. & Wilson, G. D. (1995). Proliferation measurements with flow cytometry Tpot in cancer of the uterine cervix: preliminary results. Int J Radiat Oncol Biol Phys 32: 1319–1329.
Wheeless, L. L., Coon, J. S., Cox, C., Deitch, A. D., deVere White, R. W. & Fradet, Y. (1991). Precision of DNA flow cytometry in inter-institutional analyses. Cytometry 12: 405–412.
Wheeless, L. L., Coon, J. S., Cox, C., Deitch, A. D., deVere White, R. W. & Koss, L. G. (1989). Measurement variability in DNA flow cytometry of replicate samples. Cytometry 10: 731–738.
White, R. A. & Terry, N. H. A. (1992). A quantitative method for evaluating bivariate flow cytometric data obtained using monoclonal antibodies to bromodeoxyuridine. Cytometry 13: 490–495.
White, R. A., Terry, N. H. A., Meistrich, M. L. & Calkins, D. P. (1990). An improved method for computing the potential doubling time using monoclonal antibodies to bromodeoxyuridine. Cytometry 11: 314–317.
Wilson, G. D., McNally, N. J., Dische, S., Saunders, M. I., Des Rochers, C., Lewis, A. A. & Bennett, M. H. (1988). Measurement of cell kinetics in human tumours in vivo using bromodeoxyuridine incorporation and flow cytometry. Br J Cancer 58: 423–431.
Wilson, M. S., West, C. M., Wilson, G. D., Roberts, S. A., James, R. D. & Schofield, P. F. (1993a). An assessment of the reliability and reproducibility of measurement of potential doubling times (Tpot) in human colorectal cancers. Br J Cancer 67: 754–759.
Wilson, M. S., West, C. M. L., Wilson, G. D., Roberts, S. A., James, R. D. & Schofield, P. F. (1993b). Intra-tumoural heterogeneity of tumour potential doubling times (Tpot) in colorectal cancer. Br J Cancer 68: 501–506.
Withers, H. R., Taylor, J. M. G. & Maciejewski, B. (1988). The hazard of accelerated tumour clonogen repopulation during radiotherapy. Acta Oncol 27: 131–146.
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Wilson, G., Paschoud, N., Pavy, JJ. et al. Reproducibility of measurements of potential doubling time of tumour cells in the multicentre National Cancer Institute protocol T92-0045. Br J Cancer 79, 323–332 (1999). https://doi.org/10.1038/sj.bjc.6690052
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DOI: https://doi.org/10.1038/sj.bjc.6690052
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