DNA flow cytometry of paraffin-embedded tissue.
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Animals↗
Biomedical subjects
E A Musgrove
Publications and source records attributed to E A Musgrove.
The influence of age on the DNA ploidy levels of breast tumours.
Primary tumour DNA content and estimates of cell cycle kinetic parameters were analysed by flow cytometry in 114 cases of breast cancer. Tumours were classified as: near-diploid, single aneuploid, tetraploid and greater, and multiploid (defined as having more than one aneuploid tumour cell population). No significant correlations were found between ploidy and histologic type, tumour size, lymph node involvement or receptor (oestrogen and progesterone) status. a highly significant correlation between ploidy and proliferative activity (as assessed by the percentage of cells in S phase) was observed, with near-diploid and diploid tumours being associated with a low (less than or equal to 10%) S phase fraction (P = 0.0001). A marked relationship between ploidy and patient age was also seen, with increased DNA content being associated with older patients (P = 0.025). In contrast, no patients with multiploid tumours were over 60 yr, and their age distribution was significantly different from the population as a whole (P less than 0.05), suggesting that multiploidy might be a phenomenon associated with the menopause.
Adult↗
Ploidy as a prognostic factor in ovarian cancer.
The cellular DNA content of 50 ovarian common epithelial carcinomas was determined by flow cytometry, and tumours were classified as being either diploid or aneuploid. A significant association between tumour stage and ploidy was demonstrated, with all diploid tumours being of an early stage (p less than 0.001). Forty percent of early-stage tumours (FIGO stages I and II) and all late-stage tumours (FIGO stages III and IV) were aneuploid. This heterogeneity with respect to DNA content among tumours of a similar stage may allow the identification of neoplasms with a different natural history. The proportion of S-phase cells determined by flow cytometry is a measure of cellular proliferation and may also be of prognostic significance. Diploid tumours had a median S phase of 9.8% (2.4-14.1%), while aneuploid tumours had a significantly higher S phase of 19.6% (7-24.7%; p less than 0.05). In this study there was no relationship between histological grading of invasive carcinomas and ploidy, but in view of the relatively small numbers and limited follow-up, it was not possible to perform a multivariate analysis of all known prognostic factors in ovarian cancer. Our results suggest that ploidy reflects tumour behaviour, but prolonged follow-up and increased patient accrual is necessary to assess whether the flow cytometric analysis of DNA content will provide clinically important information in ovarian cancer.
Adenocarcinoma↗
Method for analysis of cellular DNA content of paraffin-embedded pathological material using flow cytometry.
A method has been developed that allows flow cytometry to be used for measuring the cellular DNA content of paraffin-embedded human tumors. Thick (i.e., 30 micron) sections were cut from tissue blocks using a microtome and dewaxed in xylene. The sections were then rehydrated by sequentially immersing them in 100, 95, 70, and 50% ethanol before finally washing in distilled water. Single cell suspensions were then prepared by incubation in 0.5% pepsin, pH 1.5, at 37 degrees C for 30 min. The cells were counted, washed, and stained with 1 microgram/ml 4',6'-diamidino-2-phenylindole for 30 min, and DNA content was measured using an ICP 22 flow cytometer. There was a good correlation between the DNA histograms produced using this method and those obtained using unfixed tissue from the same tumor stained with ethidium bromide plus mithramycin. This method allows the retrospective study of archival material where the clinical outcome is already known, and it should, therefore, be particularly useful for determining the prognostic significance of abnormal DNA content measured by flow cytometry.
DNA, Neoplasm↗