DNA ploidy and pS2 protein expression in breast cancer

The DNA content of ductal breast carcinomas of varying histological grade was measured using static image cytometry and correlated with pS₂ expression in the tumour cells. Our study was performed on imprint of surgical biopsies of 60 women with ductal breast cancer. A statistically significant difference was observed between pS₂⁺ expression and grade of malignancy ( P <0.001). The percentage of euploid tumours significantly decreased from grade I to grade II to grade III ( P =0.01). The percentage of aneuploid tumours increased from pS₂⁺ to pS₂⁻ breast tumours ( P <0.001). These findings may be indicative of pS₂ and DNA ploidy alterations and tumour aggressiveness.     

DNA content and p53 protein expression in ductal breast cancer

DNA content and p53 protein expression in ductal breast cancer
The DNA content of 85 ductal breast cancers of different histological grades was evaluated using static cytometry and correlated with immunocytochemical expression of p53 protein in tumour cells in cytological material. A statistically significant difference was observed between p53 protein expression and grade of malignancy ( P <0.001). The percentage of euploid tumours significantly decreased from grade I through grade II to grade III tumours ( P <0.001). Clonal DNA heterogeneity was observed in 26.6% of cases analysed and was correlated with p53 protein expression ( P <0.001). These changes probably reflect genomic alterations which may affect potential malignancy of breast cancer.     

p53 Protein expression and proliferative activity in ductal breast carcinomas

The immunocytochemical expression of p53 protein and Ki-67 labelling index in tumour cells of 100 ductal breast carcinomas of different histological grade and stage was evaluated in cytological material. In order to investigate p53 expression and Ki-67 expression an avidin-extravidin immunocytochemical technique was applied to imprints. Monoclonal antibody (MoAb) DO-p53 and proliferating cell monoclonal antibody were used as primary antibodies. A statistically significant difference was observed between p53 protein expression and grade of malignancy and clinical stage (P = 0.001, P < 0.001, respectively). A statistically significant difference was also observed between Ki-67 LI and histological grade and stage of the tumours (P < 0.001, P < 0.001 correspondingly). A correlation was observed between p53 protein expression and Ki-67 LI (P < 0.001). The immunocytochemical study of p53 protein and Ki-67 expression in cytological material represents a simple method which can be applied in routine cytological laboratories for the investigation of potential malignancy of ductal breast cancer.     

Cytologic grading and DNA image cytometry of breast carcinoma on fine needle aspiration cytology smears

OBJECTIVE: To correlate the cytologic grade of breast carcinoma with DNA image cytometry (ICM) and nuclear area on fine needle aspiration cytology (FNAC) smears. STUDY DESIGN: In this prospective study, FNAC material from 28 breast carcinomas were studied for cytologic grade and DNA ICM. Breast carcinomas were classified as grade 1-3 (low to high). DNA histograms were classified by the modified Auer method. Degree of hyperploidy (DH), ploidy balance (PB) and nuclear area (NA) were measured on Feulgen-stained smears by a CAS 200 image cytometer. Cytologic grade was correlated with DNA ICM findings and NA. RESULTS: There were 3 cytologic grade 1, 13 grade 2 and 12 grade 3 breast carcinomas. Seven of eight cases of hypertetraploid aneuploidy were grade 3 tumors. All cytologic grade 1 tumors were diploid. There were significant differences in DH, PB and NA in different grades of breast carcinoma (one-way ANOVA). CONCLUSION: DNA image cytometry in combination with cytologic grading might offer additional information for the characterization of breast carcinomas diagnosed by FNAC. These observations are of particular interest with the introduction of preoperative chemotherapy.     

Comparison of breast carcinomas diagnosed in the 1980s with those diagnosed in the 1940s to 1960s.

OBJECTIVE--To find out if breast carcinomas diagnosed in the 1980s differ from those diagnosed a few decades ago. DESIGN--Retrospective comparative cohort study. SETTING--City of Turku, south western Finland. PATIENTS--439 patients with breast carcinoma diagnosed in 1945-65 with a median follow up of 27 years (95% of all those histologically diagnosed in Turku); and 370 patients with breast carcinoma diagnosed in 1980-4 with a median follow up of 6 years (94% of all those histologically diagnosed in Turku). MAIN OUTCOME MEASURES--Breast cancer incidence, mortality from breast cancer, age at diagnosis, stage of cancer, seven histological factors, DNA ploidy. RESULTS--Age adjusted incidence of breast cancer increased from 30.8/100,000 person years in 1953-7 to 62.2/100,000 in 1983-7; mortality in breast cancer increased from 16.7 to 17.2/100,000 person years. Survival of patients with stages II to IV (but not with stage I) improved. The mean age at diagnosis increased from 55.5 years in 1945-55 to 62.5 years in 1980-4 (p less than 0.0001); the proportion of patients with T0-1 carcinomas increased from 13% to 41% (p less than 0.0001) and with pN0 carcinomas from 43% to 55% (p = 0.003) from 1945-65 to 1980-4. There was no change in histological type or DNA ploidy of breast cancer, but in the 1980-4 cohort carcinomas were more often well differentiated, had lower mitotic counts and less nuclear pleomorphism, more often had a well defined tumour margin, and had less tumour necrosis. There was, however, no difference between the two cohorts in these histological characteristics except for tumour necrosis when they were compared in a multivariate log linear model at a given size of primary tumour. CONCLUSION--Improved survival with breast cancer can at least partially be explained by detection of increased numbers of small carcinomas with favourable histological characteristics.     

Image cytometry of aneuploidy, growth fraction (MoAb Ki-67) and hormone receptors (ER, PR) immunocytochemical assays in breast carcinomas

DNA nuclear content was assessed in human breast carcinomas (n = 132) using image cytometry. Optical density histograms of Feulgen stained cell imprints from fresh tissue samples, subsequently frozen for immunocytochemical assays, were determined by the SAMBA system and used for the DNA index, the ploidy balance (PB) and the proliferation index (PI) computation. The three parameters were correlated to (i) histological data (tumour grade, vascular and/or lymph node invasion) and to (ii) growth fraction (Ki67), hormone receptor antigenic sites (ER, PR) and intramedullar (bone marrow) biopsies and anti-KL1-positive epithelial cells. It was shown that 57% of breast carcinomas were aneuploid. Aneuploidy PI significantly correlated to the criteria of poor prognosis such as high tumour grade, vascular and lymphatic invasion and to increased Ki67-positive cells, and the absence of or low ER and PR. Since image cytometry is easy to handle and perfectly suitable for current diagnostic practice in pathology departments, particularly for tumour cell ploidy assessment and standardized analysis of immunostaining procedures with morphological control of the preparation, we conclude that image cytometry, as performed with the SAMBA, must be regarded as a relevant tool for prognosis evaluation and therapy guidance in individual patients.     

DNA ploidy and S-phase fraction in carcinoma of the gallbladder related to histopathology, number of gallstones and survival.

Gallstones are a risk factor for the development of gallbladder cancer. We studied DNA ploidy and cell cycle composition by flow cytometry in archival specimens from 52 gall bladder carcinomas in relation to histopathological grade, tumour stage, gallstone number and survival. 69% of the gallbladder carcinomas showed aneuploidy. All tumours with single stones (N=11) were aneuploid while only 61% of tumours with multiple stones (N=41) were aneuploid (p=0.002). DNA aneuploidy was related to increase in T-category (p=0.01), grade (p=0.02), and nuclear pleomorphism (p=0.0005). The distribution of DNA ploidy shifted from tetraploid in low stage towards triploid positions in high stage tumours (p=0.02) combined with higher S-phase values in triploid tumours (p=0.05). S-phase fraction increased during development from normal tissue to dysplasia, cancer in situ and cancer in diploid cases (p=0.0002), and further at the change from diploid to aneuploid (p=0.004). At a median cancer specific survival time of four months patients with diploid tumours had a better survival than those with aneuploid tumours (p=0.02). In multivariate analysis of the tumour characteristic, only T-category and tumour grade were independent prognostic factors.The shift from diploid to aneuploid and the further shift of ploidy within aneuploid tumours are in agreement with the concept of a clonal development of gallbladder cancer. These changes are combined with a stepwise increase in the fraction of S-phase cells. Low frequency of symptoms in single stone patients may be the reason for detection of malignancy at a late stage of tumour development.     

The prognostic significance of thymidine phosphorylase, thymidylate synthase and dihydropyrimidine dehydrogenase mRNA expressions in breast carcinomas

Thymidine phosphorylase (TP), thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) have been indicated as possible predictive markers for epithelial malignancies. All these three enzymes are actively involved in 5-FU metabolism. In this report, we investigated mRNA expression of these factors with real-time quantitative PCR in a series of 86 micro-selected breast carcinomas and 8 micro-selected tumour-adjacent normal breast epithelial specimens. Highly variable mRNA expressions of these factors were observed in both normal and cancerous samples. TP and TS mRNA expressions in breast carcinomas were elevated, but only TS mRNA expression showed a trend for statistical difference, compared with the expression in normal breast epithelial samples. Although the DPD mRNA expression range in tumours was also elevated, the average mean was reduced in tumours compared to that in normal samples. No association between mRNA expressions of TP, TS and DPD and clinicopathological features such as histological grade, tumour size, node status, S-phase fraction, ploidy, and clinical stage was found. A negative association between DPD mRNA expression and age was, however, revealed. Ten-year follow-up analysis showed no association between TP and DPD mRNA expression and clinical outcome. An high level of TS mRNA expression, however, was associated with a shorter clinical survival, indicating its potential role as a clinical marker in breast carcinoma.     

p53 protein expression and oestrogen and progesterone receptor status in invasive ductal breast carcinomas

p53 protein expression and oestrogen and progesterone receptor status in invasive ductal breast carcinomas The p53 protein expression and oestrogen and progesterone receptors status was investigated in correlation to the grade of malignancy of primary breast carcinomas. Our material constituted imprints from surgical biopsies of 75 invasive ductal breast cancer cases. The p53 protein expression was investigated immunocytologically using the monoclonal antibody p53 DO-7 (DAKO). A biochemical DCC method was applied for the detection of oestrogen and progesterone receptors for all tumours. Fifty-one percent of breast cancer cases were p53 protein positive. A statistically significant association of p53 protein expression and high tumour grade was found (chi2=23.72, d.f.=2, P < 0.001). A statistically significant association was also found between oestrogen and progesterone receptor positive cases and the grade of malignancy (P < 0.001). A negative association between p53 protein expression and oestrogen (ER) and progesterone receptors (PgR) positivity was found. From our results it appears that it is possible to distinguish from grade II tumours two subgroups of cases, one with low malignancy potential and p53 (-), ER (+), PgR (+), and another subgroup with high malignancy potential and phenotype p53 (+), ER (-), PgR (-). The last subset of patients could actually benefit from adjuvant therapy.     

Prognostic significance of DNA ploidy and proliferation rate in human astrocytic gliomas

DNA ploidy and the proliferative potential in 75 gliomas were investigated using bromodeoxyuridine labelling index (BrdUrd LI), S-phase fraction (SPF) and argyrophilic nucleolar organizer regions (AgNOR) technique. There were 53 highly malignant (AIII-AIV), and 22 low-grade (AI-AII) gliomas. One fragment of the tumour was fixed in Carnoy's solution for AgNOR test, while the other fragments were used for flow cytometric determination of the labelling index, SPF and DNA ploidy. For the BrdUrdLI, tumour samples from each patient were incubated in vitro for one hour at 37 degrees C with BrdUrd using the high pressure oxygen method. The tumours showed variability in the BrdUrdLI values, SPF and AgNOR counts/cell nucleus. The same percentage of DNA aneuploidy (55%) was found in high-grade as well as in low-grade gliomas. Univariate analysis showed that patients with grade I & II gliomas had significantly higher 3-year survival rate (p = 0.0193) than those with grade III and grade IV gliomas. Also patients with lower proliferation rate of tumours (BrdUrdLI < or =2.3% and AgNOR counts < or =2.6%/cell) had higher 3-year survival rate (p<0.03), which can be helpful in prognosis. Tumour ploidy or SPF had no influence on patients' survival (p = 0.7908). Cox multivariate analysis showed that only patients' age > 45 years and high tumour grade (III and IV) were significant unfavourable prognostic factors in terms of patients' survival.     

p53 expression measured by flow cytometry. A comparison of three monoclonal antibodies and the relationship with grade and DNA ploidy in breast cancer

The aim of this study was to quantify p53 expression by flow cytometry. A panel of three monoclonal antibodies: NCL-p53-240, NCL-p53-1801 and NCL-p53-DO7, was tested on breast cell lines and primary breast cancers. The relationships between ploidy, tumour grade and p53 expression for each antibody, were examined. Methodology was assessed using a variety of breast cell lines. Staining patterns were confirmed and the quantification technique qualified. Cytokeratin-positive cells from 58 samples obtained from patients with breast cancer were assayed for DNA content and p53 expression. p53 quantification was performed using calibrated fluorescent beads on cytokeratin-positive cells. Bloom and Richardson grading revealed 20 grade I and 38 grade II/III breast cancers. Examination of fluorescence thresholds showed a positive correlation between grade and DO7 (P=0.003) at a level of 8900 molecules, 240 (P=0.005) at a level of 2900 molecules and 1801 (P=0.005) at a level of 1850 molecules. These levels equated with 34% (DO7), 43% (240) and 43% (1801) of the samples being classified as p53-positive. Examination of ploidy revealed 23 diploid and 35 aneuploid breast cancers. Application of p53 threshold levels on diploid and aneuploid tumours showed correlation between aneuploidy and p53 expression for DO7 at a level of 9000 molecules, 240 at a level of 1900 molecules and 1801 at a level of 1800 molecules. These levels equated with 34% (DO7), 52% (240) and 52% (1801) of the samples being classified as p53-positive. We conclude that measurement of p53 by flow cytometry may be of clinical importance by indicating levels of positivity using fluorescence thresholds. p53 expression has been shown to correlate with both grade and ploidy. Flow-cytometric measurement of p53 may be a useful prognostic assay.This study was supported by the North of England Cancer Research Campaign     

Terminal uridine nick end labelling and mitosis in breast carcinoma. Correlation with tumor grade and p53 overexpression

OBJECTIVE: To study the relationship of cell death and proliferation to histologic grade and p53 expression in invasive carcinoma of the breast. STUDY DESIGN: A total of 31 cases of infiltrating duct carcinoma of the breast were randomly selected. The terminal deoxynucleotidyl-transferase-mediated dUTP nick end labelling (TUNEL) reaction and p53 immunostaining were performed on representative paraffin-embedded tissue sections. Mitotic and apoptotic indices (MI and AI) were also measured on hematoxylin-eosin-stained sections. Histologic grade of infiltrating duct carcinoma was performed with the help of the Nottingham modification of the Bloom-Richardson system. Tumor grade and p53 overexpression were correlated with MI, AI and AI detected by TUNEL. RESULTS: There were a total of 31 infiltrating duct carcinomas of the breast, of which 13 cases were grade 1 and nine cases each were grade 2 and 3. Cells with positive TUNEL showed a strong brown nuclear positivity. TUNEL showed positivity from the periphery of the nuclear margin to the central portion. AI detected by TUNEL did not correlate with tumor grade (ANOVA, P > .05). MI was significant only in grade 1 versus grade 3 and 2 versus grade 3 carcinomas (ANOVA, P < .01). The morphologic apoptotic index was significant only in grade 1 versus grade 3 carcinomas. Nine cases showed p53 overexpression, and the rest of the cases were negative for p53 immunostaining. MI, AI and TUNEL were not significantly different in p53-negative and -positive groups. Pearson's correlation coefficient showed that AI and MI were significantly related, but there was no significant relation between AI detected by TUNEL and MI. CONCLUSION: MI is still more useful than AI or AI detected by TUNEL in differentiating various grades of carcinoma of the breast.     

Reduced expression of Claudin-7 in fine needle aspirates from breast carcinomas correlate with grading and metastatic disease

OBJECTIVE: To study the immunocytochemical expression of the tight junction protein Claudin-7 in smears from breast carcinomas and correlate with grading, nodal status, locoregional and distant metastases and the cellular cohesion. METHODS: The material consisted of 52 air-dried smears from fine needle aspirates of breast carcinomas, both primary and metastatic and smears from seven benign lesions. A primary antibody to Claudin-7 was used for immunocytochemical staining. The degree of staining was recorded as negative, reduced or full, with full expression meaning equivalent to the staining pattern found in the fibroadenomas used as benign control. Staining intensity and the percentage of stained cells were evaluated. The control smears revealed a strong membrane and cytoplasmic positivity in all luminal epithelial cells. Cellular cohesion was graded as: (1) mainly cohesive groups, (2) groups and single cells and (3) mainly single cells. RESULTS: All primary and recurrent/metastatic breast lesions expressed Claudin-7. Full expression was demonstrated in 46% of the cases. Reduced expression was found in 54%. In cases with reduced expression, the percentage of stained cells were usually high, and no smear showed <50% stained tumour cells. The staining pattern was heterogeneous and always mixed membrane/cytoplasmic. Claudin-7 expression showed a significant correlation (P < 0.05) with grading, locoregional and distant metastases, nodal involvement and cellular cohesion in invasive carcinomas, but not with tumour size or subtype. CONCLUSION: Reduced expression of Claudin-7 correlated with higher tumour grade, metastatic disease, including loco-regional recurrences and with cellular discohesion.     

Nucleolar organizer regions in meningiomas. Correlation with histopathologic malignancy grading, DNA cytometry and clinical outcome

Eighty-one meningiomas (63 grade I, 9 grade II and 9 grade III) and 2 meningeal sarcomas (grade IV) were investigated by a simple one-step silver staining for nucleolar organizer region (NOR)-associated proteins (AgNOR technique) and by DNA cytometry. The number of NORs per cell and the NOR area per cell were correlated with the histopathologic grading, as were the 5c exceeding rate and the 2c deviation index (2cDI) obtained by DNA cytometry. The differences in NOR parameters were only significant (at P less than .001) between grades I and II; P was less than .05 for the 2cDI between grades I and II. No significant differences between grades II and III were found. Among recurrent tumors, the AgNOR technique revealed the proliferative potential in 8 of 11 tumors studied, whereas DNA cytometry failed to recognize malignant features in 8 of 10 tumors investigated.     

Tumour ploidy, morphometry, histological grading and clinical features in ovarian carcinoma: mutual relations

The relationship between tumour ploidy and qualitative and quantitative histopathology was assessed in a series of 95 ovarian carcinomas. 67% of the tumours were non-diploid (DNA aneuploid). 56% of the early stage (I-II) tumours were non-diploid and 81% of the tumours in advanced (III-IV) stages were aneuploid. Histological grading failed to show a clear relationship between increasing malignancy grade and ploidy. There was a close association between DNA ploidy and nuclear perimeter, area and shortest and longest nuclear diameter: the nuclei of non-diploid tumours were generally larger. Also the number of mitotic figures per square millimeter of epithelium in the microscope image (volume-corrected mitotic index, M/V-index) differed significantly between near-diploid and non-diploid tumours. Discriminant analysis showed that 74% of the learning-set tumours (67% of the test set tumours) could be correctly classified in low-ploidy and high-ploidy categories with morphometric features (nuclear perimeter, M/V-index and volume percentage of epithelium). Characteristic features of non-diploid ovarian tumours--rapid proliferation and large nuclear size--could be assessed with morphometric methods which allowed a relatively large aneuploid tumour group to be distinguished.     

乳腺癌中Vimentin与P-糖蛋白表达和其它预后指标的相关性研究

检测乳腺癌 V im entin与 P-糖蛋白的表达 ,对其相关性进行研究 ,同时与其它病理指标进行比较分析。应用免疫组织化学技术 (S- P法 )对 6 1例浸润性乳腺癌石蜡包埋组织进行检测。Vim entin阳性 2 2例 (36 .1% ) ,P-糖蛋白阳性 18例(2 9.5 % ) ,在 P-糖蛋白阳性或阴性表达组中 ,Vimentin的阳性表达率分别为 6 6 .7% (12 / 18)和 2 3.3% (10 / 43) ,两者呈显著的正相关 (P<0 .0 5 ) ;同时 ,Vimentin的表达在高病理分级病例中为 5 7.1% ,亦呈显著的正相关 (P<0 .0 5 ) ,而与淋巴结转移和 PCNA指数均无明显的相关性 (P>0 .0 5 )。在乳腺癌中 V im entin的表达可能是多药耐药性的一个表型 ,且表明肿瘤细胞分化程度较低 ;所以 Vimentin的表达提示病人预后较差     

Flow cytometric study of DNA distribution in cytopunctures of benign and malignant breast lesions

One hundred seventy-eight cytopunctures of mammary lesions were obtained for cytologic diagnosis and flow cytometric (FCM) analysis of the nuclear DNA content. All lesions were excised and evaluated histologically; 106 were carcinomas and 72 were benign lesions. The benign lesions showed a diploid DNA content, with one exception. Among the 106 carcinomas, 35 (33%) were diploid, 14 (13%) were tetraploid and 57 (54%) were aneuploid. For 79 carcinomas, the relationship between ploidy and (1) "T" and "N" of TNM staging, (2) the histologic grading of Scarff, Bloom and Richardson, (3) axillary nodal involvement, (4) the presence of estrogen and progesterone receptors, (5) age and (6) menopausal status was investigated. The percentage of aneuploidy was significantly higher (P less than .05) in grade III tumors as compared to grade I tumors. There was no significant relationship between aneuploidy and the other factors. However, a trend was observed between the lack of steroid receptors and a high probability of the tumor being aneuploid. FCM DNA analysis was also carried out on breast carcinomas obtained at surgery in 40 patients for whom FCM DNA analysis had previously been performed on breast cytopuncture specimens. The FCM DNA analyses were found to be best performed on the samples obtained by cytopuncture, which may increase the yield of tumor cells.     

磁共振成像表观扩散系数与乳腺浸润性导管癌组织学分级及预后指标的相关性研究

目的:探讨磁共振成像(MRI)表观扩散系数(ADC)与乳腺浸润性导管癌组织学分级及其预后指标的相关性。方法:收集2016年5月至2017年5月于我院就诊的并经手术病理确诊为乳腺浸润性导管癌的患者112例作为研究对象,选取患者乳腺癌组织样本作为病例组,同时选取患者对侧正常乳腺组织样本作为对照组,所有患者均行常规MRI和磁共振扩散加权成像(DW-MRI)检查,分别测量两组样本的ADC值,比较不同乳腺浸润性导管癌组织学分级与正常乳腺组织的ADC值,分析乳腺浸润性导管癌组织的ADC值与肿瘤直径大小、淋巴结转移状态、有无远处转移及雌激素受体(ER)、孕激素受体(PR)和Ki-67表达的关系,并分析ADC值与组织学分级及预后指标的相关性。结果:乳腺浸润性导管癌病理分级I级的ADC值低于对照组,病理分级II级的ADC值低于病理分级I级及对照组,病理分级III级的ADC值低于病理分级II级、I级及对照组,差异均具有统计学意义(P0.05)。乳腺浸润性导管癌患者中,肿块直径2 cm、无淋巴结转移、ER阴性、PR阴性、Ki-67阴性患者的平均ADC值均高于肿块直径≥2 cm、有淋巴结转移、ER阳性、PR阳性、Ki-67阳性患者,差异均具有统计学意义(P0.05);而有无远处转移患者之间比较差异无统计学意义(P0.05)。经Spearman秩相关分析结果显示,乳腺浸润性导管癌患者的ADC值与病理组织学分级呈现负相关关系(rs=-0.716,P=0.000);与肿块直径大小、有无淋巴结转移及ER、PR、Ki-67的表达均呈负相关(rs=-0.316、-0.545、-0.667、-0.598、-0.443,P均0.05),与有无远处转移无相关性(rs=0.091,P=0.887)。结论:乳腺浸润性导管癌的ADC值与癌组织学分级和预后相关指标存在一定相关性,可作为一种临床诊断和判断预后的重要指标,具有重要临床价值。     

Prognostic significance of DNA ploidy determined by high-resolution flow cytometry in breast carcinoma

OBJECTIVE: To assess the prognostic value of DNA ploidy in breast carcinoma and its relation to other established prognostic factors. STUDY DESIGN: We evaluated DNA ploidy in 303 breast carcinoma patients with a median follow-up of 63 months. Flow cytometry was performed on frozen tumor material, yielding histograms with narrow peaks (median coefficient of variation of 2.08). DNA ploidy pattern was classified as either diploid versus nondiploid, euploid (diploid and tetraploid) versus aneuploid or diploid/near-diploid (DNA index < 1.2) versus other, and correlated with relapse-free (RFS) and cancer-specific survival (CSS) along with tumor size, histologic grade and type, axillary lymph node involvement, menopausal and steroid receptor status, age and type of treatment. RESULTS: Seventy-one percent of tumors were DNA nondiploid (14% tetraploid and 57% aneuploid). There was a strong association between DNA ploidy and histologic grade. Histologic grade, lymph node status, tumor size and DNA ploidy (regardless of the classification used) were all significantly associated with RFS and CSS in multivariate analysis. CONCLUSION: These results suggest that DNA ploidy, at least when determined from frozen tumor tissue, is an independent prognostic factor in breast carcinoma; however, its prognostic power seems to be inferior to that of histologic grade, with which it strongly correlates.