DNA analysis by flow cytometry, response to endocrine treatment and prognosis in advanced carcinoma of the breast

117

Summary The nuclear DNA content of 351 breast carcinomas was determined by flow cytometry from paraffin-embedded tissue to assess the prognostic significance of DNA ploidy, the DNA index (DI) and the S-phase fraction (SPF). The minimum follow-up of the patients was 22 years, and they were all from a defined urban population. DNA ploidy correlated with histological type and grade, mitotic count and nuclear pleomorphism (P<0.0001), and also with axillary nodal status (P=0.0005), tumour necrosis (P=0.001), primary tumour size (P = 0.03), menopausal status (P = 0.004) and the presence of distant metastases at the time of the diagnosis (P = 0.04). Survival corrected for intercurrent deaths of the patients with a diploid tumour was better than that of the patients with a non-diploid tumour (P = 0.0001, 48% vs 28% at 25 years). SPF Leatham & Brooks, 1987; Slamon et al., 1987).The nuclear DNA content determined by flow cytometry has recently emerged as a new prognostic factor in breast cancer. The technique has gained some popularity, since flow cytometric histograms are rapid to produce and often easy to interpret. It has been suggested that breast carcinomas with an abnormal (non-diploid or aneuploid) nuclear DNA content are associated with less favourable prognosis than carcinomas with a normal (diploid) DNA content (Hedley et al., 1984(Hedley et al., , 1987Coulson et al., 1984;Ewers et al., 1984;Thorud et al., 1986;Baildam et al., 1987;Cornelisse et al., 1987;Kallioniemi et al., 1987;Stal et al., 1989). Similarly, the DNA index (DI, the relative DNA content of an aneuploid stemline of cells as compared with diploid cells) and the percentage of S-phase cells in the DNA histogram (S-phase fraction, SPF) have been found by some to be of prognostic significance (Dowle et al., 1987;Hedley et al., 1987;Kallioniemi et al., 1988;Stal et al., 1989;McDivitt et al., 1986;Klintenberg et al., 1986). However, the prognostic value of nuclear DNA content analysis is still unsettled. Some authors report DNA content to be the most important prognostic variable in breast cancer, and to have independent prognostic value in a multivariate analysis , whereas others find it to have prognostic value only in a univariate analysis (Stal et al., 1989;Hedley et al., 1984), and yet a few fail to find any prognostic significance at all when overall survival is concerned (van der Linden et al., 1989;Uyterlinde et al., 1988;Owainati et al., 1987

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confidence: 99%

The prognostic significance of nuclear DNA content in invasive breast cancer – a study with long-term follow-up

Toikkanen

Joensuu²,

Klemi³

1989

117

“…Following the paper of Hedley et al (1983) describing flow cytometric analysis of DNA using formalin-fixed and paraffin-embedded tissues, numerous studies have been published confirming the application of their method to archival material (Friedlander et al, 1984;Hiddemann et al, 1984;Coon et al, 1986;Douglas et al, 1986;Schmidt et al, 1986;Baildam et al, 1987). Until then, information on the DNA content of tumours was obtained using either Feulgen microspectrophotometry, which is a painfully slow procedure allowing only a small number of cells to be examined, or flow cytometry, which needed fresh unfixed tissue or karyotyping (Atkin, 1972;Mann & Yates, 1979).…”

mentioning

confidence: 99%

Prognostic relevance of DNA content in childhood renal tumours

Kumar

Marsden²,

Cowan³

et al. 1989

Summary The DNA content of paraffin embedded tumour specimens from 100 children with kidney tumours was studied by flow cytometry. Data of adequate quality were obtained from 93 cases comprising 67 Wilms' tumours with a favourable histology (FH), 12 Wilms' tumours with unfavourable histology (UH) (pleomorphic), 8 bone-metastasising renal tumours of childhood (BMRTC) and 6 rhabdoid renal tumours. Only 4.5% FH compared with 75% UH Wilms' were aneuploid (P<0.001). Although BMRTC and rhabdoid tumours are associated with poor prognosis, there were no examples of aneuploidy in these tumours. The proliferation index was found to be of no prognostic value. Staging and ploidy were not correlated with each other in any of the various histological types of renal tumours studied.

“…There is good evidence from both static and flow cytometric DNA analysis of paraffin embedded material or freshly excised tissue that patients with DNA diploid tumours are likely to have a better prognosis than their DNA aneuploid counterparts (Auer et al, 1980;Baildam et al, 1987;Kallioniemi et al, 1987a, b). We have recently reported that sufficient cellular material for meaningful flow cytometric DNA analysis can be obtained from the majority of routine diagnostic FNAs (Levack et al, 1987).…”

Section: Discussionmentioning

confidence: 99%

“…While there was no variation between sequential FNAs in four patients, marked differences in the DNA profile were observed in the remaining nine patients undergoing treatment, the changes not necessarily being associated with clinical response to therapy. It is concluded that the monitoring of cellular changes by DNA analysis of sequential FNAs may be complex and subject to problems associated with heterogeneity.Flow cytometric DNA analysis can provide useful information on the cellular characteristics of breast tumours (Auer et al, 1980;Baildam et al, 1987;Cornelisse et al, 1987;Dowle et al, 1987; Kallioniemi et al, 1987a, b;Owainati et al, 1987). Recently, we demonstrated that fine needle aspiration of most breast cancers can provide sufficient cellular material for meaningful analysis of DNA profiles using flow cytometry (Levack et al, 1987).…”

mentioning

confidence: 99%

“…Flow cytometric DNA analysis can provide useful information on the cellular characteristics of breast tumours (Auer et al, 1980;Baildam et al, 1987;Cornelisse et al, 1987;Dowle et al, 1987; Kallioniemi et al, 1987a, b;Owainati et al, 1987). Recently, we demonstrated that fine needle aspiration of most breast cancers can provide sufficient cellular material for meaningful analysis of DNA profiles using flow cytometry (Levack et al, 1987).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Variations associated with the DNA analysis of multiple fine needle aspirates obtained from breast cancer patients

Mullen

Miller²

1989

Summary The present study was carried out to determine the variation in DNA content between multiple fine needle aspirates (FNA) of the same tumour from patients with breast cancer. Analysis of different aliquots of the same FNA showed good reproducibility in terms of cell cycle distribution and DNA index. Duplicate FNAs taken from different sites in nine of 11 excised tumours showed similar reproducibility. However, two of the aneuploid tumours displayed substantial variations in the distribution of cell populations between the duplicate samples. Sequential FNAs with no intervening therapy were obtained from the same tumour in 17 patients; one at the time of diagnosis and the other at biopsy 1-3 weeks later. Only five cases showed no variation between the sequential FNAs; the remaining 12 displayed different DNA profiles. A further 13 patients were studied before and during systemic therapy. While there was no variation between sequential FNAs in four patients, marked differences in the DNA profile were observed in the remaining nine patients undergoing treatment, the changes not necessarily being associated with clinical response to therapy. It is concluded that the monitoring of cellular changes by DNA analysis of sequential FNAs may be complex and subject to problems associated with heterogeneity.Flow cytometric DNA analysis can provide useful information on the cellular characteristics of breast tumours (Auer et al., 1980;Baildam et al., 1987;Cornelisse et al., 1987;Dowle et al., 1987; Kallioniemi et al., 1987a, b;Owainati et al., 1987). Recently, we demonstrated that fine needle aspiration of most breast cancers can provide sufficient cellular material for meaningful analysis of DNA profiles using flow cytometry (Levack et al., 1987). This use of flow cytometry in the analysis of cellular material from FNAs has potentially wide applications. For example, it may be possible to monitor the effects of treatment by analysing cellular characteristics of sequential tumour FNAs.In this respect it is important to show that analysis of FNAs provides reproducible results. The aim of the present paper was to assess variation in DNA profiles by studying multiple fine needle aspirates taken simultaneously and sequentially from breast tumours. Patients and methods PatientsTumour fine needle aspirates (FNAs) were obtained from patients attending the Professorial Breast Clinics of the University Department of Surgery, Edinburgh. All tumours were shown histologically to be breast cancers and malignant cells were cytologically confirmed to be present in all aspirates. AspiratesThe method of aspiration was that described by Zajicek (1965) and used a 23 gauge needle attached to a 10 ml syringe. After removing an aliquot for cytological confirmation of malignancy (Dixon et al., 1984), the remainder of the aspirate was expelled into 200 dp of citrate buffer and stored at -40°C until analysis (Vindel0v et al., 1983a, b values falling within the 'S' phase fraction of diploid tumour cells.Briefly, frozen cell suspensions were tha...