Quantitation by flow cytofluorometry of response of tumours of the uterine cervix to radiotherapy

Abstract

We are presently involved in a project to investigate the use of flow cytofluorometry in assessing, by means of serial biopsies, the response to radiotherapy of tumours of the uterine cervix. This technique enables changes in the DNA content profiles and content of proliferating cells in the tumour biopsies to be determined at intervals as therapy progresses. A means has been devised to quantitate the content of hyperdiploid cells, hypertetraploid cells, and dead and dying cells, by computer analysis of DNA vs. RNA scattergrams obtained by flow cytofluorometric analysis. Plotting of these parameters for the serial biopsies vs. time since start of tumour irradiation presents a graphical indication of the response of the tumour to irradiation. Over 100 patients (stage Ib and IIa) have been followed during intracavitary (Cathetron) therapy with parallel flow cytofluorometric analysis and histopathological assessment of serial biopsies. Patterns are beginning to emerge from these analyses which promise to indicate within 14-21 days and sometimes earlier the extent of radioresponsiveness of the tumour. These patterns may also be of assistance in planning modified dose fractionation schedules to obtain improved therapeutic ratios.

Heterogeneity of colorectal adenocarcinomas evaluated by flow cytometry and histopathology

“…It was found that 36 tumours (72%) would have been correctly assigned on the basis of any one sample whereas 14 tumours (28%) could have been incorrectly designated since one of the samples fell into a different ploidy group from the others. If the tumours were further subdivided into three subgroups (<10%, 10-30%, >30% of Cell death Previous studies have confirmed the validity of using ethidium bromide staining to determine the level fo cell death in vivo and in vitro (Dyson et al, 1984a(Dyson et al, , 1984b(Dyson et al, , 1984c. The range of values found in the various tumours (8-92%) was the greatest of any of the areas quantitated ( Figure 4).…”

“…Cells were again collected by centrifugation and resuspended in 2-4 ml of PBS (50%) and HBSS (50%); an equal volume of the same medium was then added containing 15,uM acridine orange (AO) and the sample was placed in the flow cytometer and the sample flow started and allowed to equilibriate. Figure 2 and were modified from our previous studies (Dyson et al, 1984a(Dyson et al, , 1984b(Dyson et al, , 1984c to allow for the wider range of DNA content seen in these tumours. Table IV.…”

“…For these reasons we have employed in this study a combination of a newly developed flow cytometric technique (Dyson et al, 1984a;1984b;1984c) with conventional histopathological assessments of primary and metastatic colorectal adenocarcinomas. We have attempted to determine the extent of tumour heterogeneity and the proportion of dying cells to assess their influence on tumour staging and grading.…”

The effect of sodium butyrate on the growth characteristics of human cervix tumour cells

“…The cervix tumour cell lines employed in this study were established in primary culture from cervix biopsy tissue taken routinely during radiotherapy at Cookridge Hospital (Dyson et al, 1984a). Six cell lines were used: 754, 612, 995, 090, 329, 708.…”

“…The cervix tumour cell lines employed in this study were established in primary culture from cervix biopsy tissue taken routinely during radiotherapy at Cookridge Hospital (Dyson et al, 1984a). Six cell lines were used: 754, 612, 995, 090, 329, 708. These were maintained as monolayer cultures from which multicellular aggregates were obtained as required by using the method of Sutherland and Durand (1976).…”