Grade Invasive Ductal Carcinoma Research Articles

Abnormal expression of Nek2 and β-catenin in breast carcinoma: clinicopathological correlations

NIMA-related kinase 2 (Nek2) and β-catenin are important centrosome regulatory factors. The aim of this study was to detect the possible disparity in their expression among normal breast tissue, invasive ductal carcinoma (IDC), concomitant ductal carcinoma in situ (DCIS), and pure DCIS, and to explore its correlation with clinicopathological factors. We used immunohistochemistry to detect protein expression of Nek2 and β-catenin in breast cancer tissues from 60 cases of pure DCIS, 348 cases of IDC and 137 cases of concomitant DCIS with that in normal breast tissues from the same 137 concomitant DCIS patients as controls. As compared with normal tissue, expression of Nek2 and β-catenin in the cytoplasm was significantly increased in IDC and DCIS (P < 0.05), and variation in expression was also observed in different grades of IDC (P < 0.01). Also, cytoplasmic expression of Nek2 and and of β-catenin were correlated with each other in IDC and DCIS (P < 0.01). In addition, they were both related to Ki67 immunoreactivity (P < 0.05). Furthermore, our study also revealed a correlation between their expression and some clinicopathological factors. We found that Nek2 cytoplasmic expression was associated with grade and tumour size (P < 0.01) in IDC, whereas β-catenin cytomembrane expression showed significant variation with grades, TNM stages, lymphoid node status, oestrogen receptor status, and molecular subtype (P < 0.05); a difference in expression was also observed between IDC and DCIS (P < 0.05). Also, β-catenin cytoplasmic expression was associated with TNM stage (P < 0.05). Expression of Nek2 at the mRNA level was detected in 50 pairs of breast cancer specimens and matched normal tissues by reverse transcriptase polymerase chain reaction, and the result showed increased expression in IDC. This study suggests that abnormal expression of Nek2 and β-catenin might be one of the mechanisms of tumorigenesis, especially of abnormal tumour proliferation. They may represent new potential targets for therapeutic intervention.

Early stages of clear cell (glycogen rich cell) invasive ductal carcinoma (IDC) of the breast: Good or bad prognosis?

e11053 Background: The presence of clear cells (glycogen rich cells) in invasive ductal carcinoma of the breast is infrequent. The main objective of the present paper is to analyze the frequency of this tumor subtype in our patient group. Methods: A search of the IOHM breast cancer tumors registry was carried out. In this database all 1680 tumors were reviewed by two pathologists and classified according to the USA Association of Directors of Anatomic and Surgical Pathology’s recommendations. The entire cohort was diagnosed, treated and followed at the IOHM. Only those with early stages of clear cell (glycogen rich cell) invasive ductal carcinoma of the breast were included in this paper. Results: Between January 1998 and June 2008, 945 patients with IDC and early breast cancer were identified. 17 (1,8%) of those cases presented clear cell carcinomas, and are the subject of this report. Population Characteristics: Median Age 60 years (35-88); Menopause YES/NO 11/6; High Tumor Grade 15; Predictive Factors: ER (+):=15/17, PR (+)= 13/17, Her2 (+) 2/6 (Undetermined 11 cases), Diagnosis Stage (I=6 pts, II=5 pts, III=6 pts); Surgery: Mastectomy= 3 pts, Tumorectomy= 14pts, Radiotherapy=16 pts, Adjuvant treatment: 17 pts (Only Hormonotherapy: 7 pts, Only Chemotherapy: 5 pts, Chemotherapy and hormonotherapy: 5 pts) With a median Follow Up of 97 months (27-146), twelve patients are alive and free of disease (Stage I: 6 pts, Stage II : 4 pts, Stage III: 2 pts), Four patients have recurred (Stage II: 1 pt, Stage III: 3 pts) and four have died : 3 due to the tumor and 1 due to other causes, with a global median survival of 92 months (range 16-146). In each stage, the median survival is Stage I: 86 months (44-145), Stage II: 114 months (30-129), Stage III: 69 months (16-106). Conclusions: 1) In this cohort 1.8% of patients with early IDC breast cancer possess clear cells associated with the tumor. 2) Although the number of patients is low, the evolution was better than that observed in patients with High Tumor Grade IDC at similar stages. 3) It appears that the presence of glycogen rich cells is not a factor leading to a bad prognosis.

Abstract 2954: The transient receptor potential V6 (TRPV6) calcium channel in breast cancer cell lines and clinical breast cancer tissues

Abstract TRPV6 is an important calcium channel, which is highly selective for Ca2+ and has high basal activity. In prostate tumors TRPV6 over-expression is linked to enhanced invasiveness and proliferation. While some studies have demonstrated that TRPV6 is over-expressed in clinical breast cancer samples and studies from our laboratory have shown that TRPV6 mRNA is up-regulated in a sub-set of breast cancer cell lines, a link between TRPV6 expression and breast cancer aggressiveness has not been demonstrated. Furthermore, the mechanism for the over-expression of TRPV6 in breast cancer is unknown. To explore TRPV6 gene amplification as a possible mechanism for over-expression, we used a real-time PCR copy number assay and identified a sub-set of breast cancer cells lines that exhibited increased TRPV6 gene copy number. To assess the clinical relevance of this finding, we initially used a high throughout, clinically driven approach to examine TRPV6 gene amplification in breast tumors. Our findings showed that the mechanism of TRPV6 over-expression is due to, at least in part, gene amplification and that gene amplification may be pronounced in breast cancer subtypes with a poor prognosis. In clinical breast tumor samples, microarray based comparative genomic hybridization (aCGH) showed that the TRPV6 gene region (7q34) is gained in 0/16 invasive lobular carcinomas and in 11/35 high grade invasive ductal carcinomas. When stratified by estrogen receptor status, 7/13 estrogen receptor negative, high grade invasive ductal carcinomas harbored a gain in this region. To determine whether the copy number increase observed at the gene region 7q34 specifically corresponded with amplification of the TRPV6 gene, we performed chromogenic in situ hybridization (CISH) using a probe specific for the TRPV6 gene and a breast cancer tissue microarray. Our findings suggest that a sub-set of breast cancers may be characterized by TRPV6 gene amplification. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 2954. doi:10.1158/1538-7445.AM2011-2954

The Cell of Origin of BRCA1 Mutation-associated Breast Cancer: A Cautionary Tale of Gene Expression Profiling

Breast tumours are highly heterogeneous with several distinct sub-types recognised according to their histological and molecular features. The biological basis for this heterogeneity is largely unknown, although there are some distinct phenotype-genotype correlations. These include BRCA1 mutation-associated breast cancers, which are typically high grade invasive ductal carcinomas of no special type (IDC-NSTs) with pushing margins that do not express estrogen receptor (ER), progesterone receptor (PR) or the HER2 receptor tyrosine kinase ('triple negative'). Gene expression analysis of these tumours has grouped them with so called 'basal-like' breast cancers and this, together with evidence that knock-down of BRCA1 in vitro blocked luminal differentiation, led to speculation that these tumours arose from the normal basal stem cells within the mammary gland. Recently, however, human breast tissue from BRCA1 mutation carriers was shown to contain an expanded population of luminal progenitor cells which have increased in vitro clonogenic ability. In the mouse, targeted deletion of Brca1 in luminal ER negative progenitors resulted in the formation of mammary tumours which phenocopied human BRCA1 breast tumour pathology, while the deletion of Brca1 in basal stem cells resulted in the formation of tumours which neither resembled human BRCA1 tumours or sporadic basal-like breast tumours. Importantly, however, both sets of mouse tumours were classified as 'basal-like' by methods used for human tumour classification based on gene expression profiles. This demonstrates that, as it stands, expression profiling is poor at distinguishing tumour histological subtypes and is also a poor guide to the cell of tumour origin. These human and rodent studies support an origin of BRCA1-mutation associated breast cancer (and indeed of the majority of sporadic basal-like breast cancers) in a luminal ER negative mammary epithelial progenitor. This is a key finding, as identification of the cells of origin in breast cancer subtypes makes possible the identification of key processes associated with initiation, progression and maintenance of each tumour subtype, the development of novel targeted therapies and, potentially, of new preventative approaches in high risk groups.

Abstract P6-01-04: Ductal Carcinoma In-Situ Associated with Triple-Negative Invasive Breast Carcinoma Is Often Immunohistochemically Distinct from Invasive Component

Abstract Background: Gene expression profiling studies have identified a sub-set of breast carcinoma known as basal-like carcinoma, which characteristically do not express estrogen receptor (ER), progesterone receptor (PR), or HER2neu (HER2). These triple negative invasive ductal carcinomas (TN-IDC) are associated with an aggressive clinical course and poor prognosis. Little information exists in the literature concerning ductal carcinoma in-situ (DCIS) associated with TN-IDC. We evaluated the frequency, histologic features, and receptor status of DCIS associated with TN-IDC. Design: From our institutional archives between July 2006 and August 2009, we retrieved all cases of intermediate and high grade invasive ductal breast carcinoma. Based on the immunoprofiling, invasive carcinomas were categorized into either TN-IDC or non triple-negative invasive carcinoma (non-TN-IDC). Immunohistochemistry for ER, PR, and HER2 was performed on the DCIS of the TN-IDC group and on some available cases from the non-TN-IDC group. The frequency, receptor status, and nuclear grade of DCIS associated with both groups were noted. Statistical analysis was performed using Fisher Exact Test. Results: Among a total of 224 cases, 56 (25%) were classified as TN-IDC and 168 (75%) as non-TN-IDC. DCIS was seen in 28 (50%) TN-IDC cases and in 150 (89.3%) non-TN-IDC cases (P&amp;lt;0.0001). Sufficient tumor was available for immunohistochemical staining in 23 of 28 cases with DCIS in the TN-IDC group. Within the 23 cases, the DCIS were triple negative in 15 cases (65%). Of the remaining 8 cases (35%), 4 were positive for ER and PR, but negative for HER2, while the other 4 cases were negative for ER and PR but positive for HER2. Ninety-six cases in the non-TN-IDC group had available immunostains for review, 7 of which (7.3%) displayed a different receptor status from their invasive counterparts (p=0.0016, compared with TN-IDC). Two cases of non-TN-IDC showed an associated TN-DCIS. Of the 28 cases of TN-IDC associated with DCIS, 4 (14.3%) showed lower nuclear grade in DCIS than in its invasive counterpart, compared with 22 of 150 (14.7%) non-TN-IDC cases (no significant difference). One TN-IDC case displayed two distinct morphologic patterns of DCIS; neither pattern was triple negative. Conclusion: Our study demonstrated the presence of DCIS in a considerable proportion of TN-IDC although at a significantly lower frequency than in non-TN-IDC. DCIS associated with TN-IDC was significantly more likely to display a different immunoprofile from its invasive counterpart than DCIS associated with non-TN-IDC. Citation Information: Cancer Res 2010;70(24 Suppl):Abstract nr P6-01-04.

Fourier Transform Infrared Spectroscopic Analysis of Breast Cancer Tissues; Identifying Differences between Normal Breast, Invasive Ductal Carcinoma, and Ductal Carcinoma In Situ of the Breast

A nondestructive method employing Fourier transform infrared (FTIR) microspectroscopy coupled with attenuated total reflectance (ATR) objective for the analysis of histopathological specimens is described. Malignant breast tissue specimens have been analyzed to demonstrate the hypothesis that chemical changes taking place in biological tissue can be reliably and reproducibly identified. This study is the first report to elucidate clear spectral differences between different ductal carcinoma in situ (DCIS) grades. Sixty individual cases of breast carcinoma including DCIS and invasive ductal carcinoma (IDC) and seven cases of normal breast tissues were studied using the FTIR-ATR spectroscopic technique. FTIR analysis of tissue sections has provided distinct spectra that can be used to distinguish between the nuclear grades of DCIS and IDC of the breast. It was concluded that FTIR could objectively and reproducibly discriminate between DCIS and IDC grades without sample destruction. In the future, applications of FTIR approaches should become feasible in the nondestructive express classification of grades and diagnosis of breast carcinoma.

Hypoxia and metabolic phenotypes during breast carcinogenesis: expression of HIF-1α, GLUT1, and CAIX

Hypoxia and acidosis are microenvironmental selection forces during somatic evolution in breast carcinogenesis. The effect of cobalt chloride (CoCl(2))-induced hypoxia on the expression of hypoxia-inducible factor (HIF)-1alpha, glucose transporter 1 (GLUT1), and carbonic anhydrase IX (CAIX) was assessed in breast cancer cells derived from primary sites (HCC1395 and HCC1937) and metastatic sites (MCF-7 and MDA-MB-231) by reverse transcriptase-polymerase chain reaction and immunoblotting. We analyzed these proteins' expression in tissue samples from normal breast tissue, usual ductal hyperplasia (DH), atypical ductal hyperplasia (ADH), ductal carcinoma in situ (DCIS), and invasive ductal carcinoma (IDC) using immunohistochemistry. CAIX mRNA was expressed constitutively in MDA-MB-231 cells but not in the other three cell lines. CAIX mRNA expression was increased after CoCl(2)-induced hypoxia in all four breast cancer cell lines. The expression of HIF-1alpha and GLUT1 proteins was increased after CoCl(2)-induced hypoxia in all breast cancer cell lines tested. Hypoxia significantly increased CAIX protein expression in primary cancer cells but not in metastatic ones. HIF-1alpha was not expressed in benign breast tissue, whereas it was significantly expressed in DH, ADH, DCIS, and IDC (p < 0.001). GLUT1 and CAIX were expressed only in DCIS (56.8% and 25.0%) and IDC (44.1% and 30.5%), with higher expression in high grade DCIS than low/intermediate grade DCIS (79.2% vs. 30.0%, p = 0.001 and 37.5% vs. 10.0%, p = 0.036, respectively). High CAIX expression was significantly associated with poor histological grade of IDC (p = 0.005). During breast carcinogenesis, the role of HIF-1alpha changes from response to proliferation to tumor progression. GLUT1 expression (glycolytic phenotype) and CAIX expression (acid-resistant phenotype) may result in a powerful adaptive advantage and represent an aggressive phenotype.

Impact of Pathological Characteristics on Local Relapse After Breast-Conserving Therapy: A Subgroup Analysis of the EORTC Boost Versus No Boost Trial

To investigate the long-term impact of pathologic characteristics and an extra boost dose of 16 Gy on local relapse, for stage I and II invasive breast cancer patients treated with breast conserving therapy (BCT). In the European Organisation for Research and Treatment of Cancer boost versus no boost trial, after whole breast irradiation, patients with microscopically complete excision of invasive tumor, were randomly assigned to receive or not an extra boost dose of 16 Gy. For a subset of 1,616 patients central pathology review was performed. The 10-year cumulative risk of local breast cancer relapse as a first event was not significantly influenced if the margin was scored negative, close or positive for invasive tumor or ductal carcinoma in situ according to central pathology review (log-rank P = .45 and P = .57, respectively). In multivariate analysis, high-grade invasive ductal carcinoma was associated with an increased risk of local relapse (P = .026; hazard ratio [HR], 1.67), as was age younger than 50 years (P < .0001; HR, 2.38). The boost dose of 16 Gy significantly reduced the local relapse rate (P = .0006; HR, 0.47). For patients younger than 50 years old and in patients with high grade invasive ductal carcinoma, the boost dose reduced the local relapse from 19.4% to 11.4% (P = .0046; HR, 0.51) and from 18.9% to 8.6% (P = .01; HR, 0.42), respectively. Young age and high-grade invasive ductal cancer were the most important risk factors for local relapse, while margin status had no significant influence. A boost dose of 16 Gy significantly reduced the negative effects of both young age and high-grade invasive cancer.

5154 Triple-negative high grade invasive ductal breast carcinomas are biologically heterogeneous: differences between the basal and non-basal subtypes

5154 Triple-negative high grade invasive ductal breast carcinomas are biologically heterogeneous: differences between the basal and non-basal subtypes

Cox-2 Expression in Malignant Breast Tumors

Purpose: Cox-2 expression in breast carcinoma has been reported to be related to angiogenesis, lymph node metastasis and Her-2 expression. The aim of this study is to evaluate the clinicopathologic significance of Cox-2 expression in the invasive ductal carcinomas (IDC) and intraductal carcinomas (DCIS) of the breast. Methods: The materials were 353 IDC cases and 82 DCIS cases. Immunohistochemical stain for Cox-2 was interpreted as 1＋ (weak & focal) and 2＋ (diffuse), and the relationships between Cox-2 and ER, PR, Her-2, p53, Ki-67 and bcl-2 expressions were analyzed. Results: There was no significant difference of Cox-2 expression between IDC (148/353, 41.9%) and DCIS (38/82, 46.3%). Cox-2 (2＋) expression was more frequent in low grade than intermediate and high grade IDC, but the difference was not significant statistically (P=0.0833), and there were no significant differences of Cox-2 expression according to age, tumor size, nuclear grade, lymph node metastasis in IDC and DCIS cases. In IDC cases, Cox-2 (1＋ and 2＋) expression showed positive relationships with p53 (＋) and more than 10% of Ki-67 labeling index (P=0.0029, P=0.0015), and revealed tendencies of positive relationships with ER (＋) and bcl-2 (＋) (P=0.0750, P=0.0776). However, no significant relationship between Cox-2 and Her-2 expressions was recognized. In DCIS cases, Cox-2 (2＋) expression rate was increased in cases showing negative for Her-2 (P=0.0092) and positive for bcl-2 (P=0.0486). Conclusion: Cox-2 expression seems to be involved in the development of breast carcinomas, but not related to the invasiveness. Cox-2 expression, especially 2＋, in the DCIS cases suggest a possibility of less aggressive biological behavior.

Histopathological characteristics of breast carcinomas at the Olabisi Onabanjo University Teaching Hospital, Sagamu Ogun State, Nigeria

This study examines the pathological grade as well as the estrogen progesterone receptor (ERPR) status in breast cancer materials from Olabisi Onabanjo University Teaching Hospital, Sagamu, Nigeria. All histopathologically diagnosed tissue samples from the department of Histopathology of the Olabisi Onabanjo University Teaching Hospital, Sagamu seen from January 2003 to December 2004 were examined and graded using the Elston and Ellis Modification of the Scarff-Bloom –Richardson grading system1 and stained for their estrogen/progesterone receptor status. Forty- seven cases of breast carcinoma were seen during the period under study. Of these, 44(93.7%) were invasive ductal carcinoma, with majority being poorly differentiated. Their ages ranged from 25-75 years with majority in the age group 40-59years. The mean age was 47.3 years. Of the 27 cases that had their ERPR status analyzed, 21 cases (77.8%) were ERPR negative, while only 3 (11.1%) were ERPR positive. This study shows that there is a predominance of high grade invasive ductal carcinomas that are likely to be ERPR negative. These features suggest a biologically aggressive form of breast cancer in Nigeria with the possibility of poor response to hormonal therapy. Nigerian Journal of Health and Biomedical Sciences Vol. 7 (1) 2008 pp. 23-26

Molecular profiling pleomorphic lobular carcinomas of the breast: evidence for a common molecular genetic pathway with classic lobular carcinomas

Pleomorphic lobular carcinomas (PLC) of the breast display histological features associated with classic invasive lobular carcinoma (ILC), yet they also exhibit more conspicuous nuclear atypia and pleomorphism, and an aggressive clinical behaviour. From a breast cancer progression perspective, it is unclear whether PLC is a variant of ILC or is a high-grade invasive ductal carcinoma (IDC) that has lost E-cadherin. The molecular features of 26 PLC were studied using immunohistochemistry [oestrogen receptor (ER), progesterone receptor (PR), HER2, p53 and E-cadherin], 0.9 Mb resolution, microarray-based comparative genomic hybridization (aCGH), fluorescent (FISH) and chromogenic (CISH) in situ hybridization and loss of heterozygosity. Comparative analysis was performed with aCGH data from PLC with classic ILC (16 cases) and high grade IDC (35 cases). PLCs were frequently ER- and PR-positive, E-cadherin-negative and occasionally HER2- and p53-positive. Recurrent copy number changes identified by aCGH included gains on 1q, 8q, 11q, 12q, 16p and 17q and losses on 8p, 11q, 13q, 16q and Xq. Highly recurrent 1q+ (100% of cases), 16p+ (93%), 11q- (53%) and 16q- (93%) and evidence of the der(1;16)/der(16)t(1;16) rearrangement, as detected by FISH, suggested that PLC had a 'lobular genotype'. Focal amplifications were evident at 8p12-p11, 8q24, 11q13.1-q14.1, 12q14, 17q12 and 20q13. Loss of BRCA2 was detected in 40% of PLC by LOH. Comparative analysis of aCGH data suggested the molecular features of PLC (ER/PR-positive, E-cadherin-negative, 1q+, 11q(-), 16p+ and 16q(-)) were more closely related to those of ILC than IDC, implicating an overlapping developmental pathway for these lobular tumour types. Molecular alterations found in PLC that are more typical of high-grade IDC than ILC (p53 and HER2 positivity, 8q+, 17q24-q25+, 13q(-) and amplification of 8q24, 12q14, 17q12 and 20q13) are likely to drive the high-grade and more aggressive biology of PLC.

DCIS with close or focally involved margins following breast-conserving surgery (BCS): reexcision or radiotherapy with boost?

The aim of this study was to determine the presence and histological grade of prominent ductal carcinoma in situ (DCIS) and the relationship between the histological grade of DCIS and the grade of invasive ductal carcinoma of the breast to local recurrence and survival. We have analysed 175 patients with stage I and II invasive ductal breast carcinoma who underwent breast-conserving therapy in the period 1987–1994. Patients were divided into those with carcinomas showing prominent DCIS (74 patients), and those with tumours without a DCIS component (101 patients). Ten patients developed recurrences and were treated by hormonal and/or chemotherapy. Local recurrence was observed in 2 (2%) patients without, and in 8 (10.8%) patients with prominent DCIS (χ2=12.954, P<0.005). There was a highly significant correlation between DCIS type and the histological grade of the infiltrating carcinoma (χ2=73.77, P<0.001). Our results suggest that the patients with poorly differentiated DCIS are significantly more likely to develop recurrence than patients with intermediately or well-differentiated DCIS.

Detection of Epstein-Barr virus in breast cancers with lymphoid stroma

the purpose of our work is to detect Epstein-Barr virus (EBV) in 2 types of breast cancer: medullary carcinoma and high grade invasive ductal carcinoma with lymphoid stroma. we proceeded to a retrospective study of 18 medullary carcinoma and 18 high grade invasive ductal carcinoma with lymphoid stroma. The detection of the virus was carried out by immunohistochemistry with anti-LMP2 antibody and by hybridization in situ by oligonucleotides EBER1 and EBER1. LMP1 as well as hybridization in situ were positive in 5 tumors (3 medullary carcinoma and 2 high grade invasive ductal carcinoma with lymphoid stroma). positivity was observed in tumor cells and neither in epithelial non tumoral ones nor in lymphoid cells. during numerous years, correlations between the replication of EBV and the appearance of a malignant phenotype were limited to nasopharyngeal carcinoma and to lymphoid cells. A controversy regarding the association of EBV with breast cancers has recently been reported in the literature. This cancer being very frequent, the involvement of EBV even in a small proportion of breast cancers could have important implications. Our results suggest a possible implication of EBV in these tumours but other studies are necessary.

Bilateral synchronous breast carcinomas followed by a metastasis to the gallbladder: a case report

BackgroundBreast cancer is usually associated with metastases to lungs, bones and liver. Breast carcinoma metastasizing to the gallbladder is very rare.Case presentationA 59-year-old woman presented with bilateral synchronous breast lesions. A palpable, retroareolar solid lesion of diameter equal to 5 cm was present in the right breast, and a newly developed, non-palpable lesion with microcalcifications (diameter equal to 0.7 cm) was present in the upper outer quadrant of the left breast. Modified radical mastectomy was performed on the right breast and lumpectomy after hook-wire localization was performed on the left breast, combined with lymph node dissection in both sides. The pathological examination revealed invasive lobular carcinoma grade II in the right breast and invasive ductal carcinoma grade I in the left breast. Chemotherapy, radiation therapy, trastuzumab and letrozole were appropriately administered. At her 18-month follow-up, the patient was free of symptoms; the imaging tests (chest CT, abdominal U/S, bone scan), biochemical tests, blood cell count and tumor markers were also normal. At the 20th month after surgery however, the patient developed symptoms of cholecystitis and underwent cholecystectomy. The histopathological examination revealed metastasis of the lobular carcinoma to the gallbladder.ConclusionThis extremely rare case confirms on a single patient the results of large series having demonstrated the preferential metastasis of lobular breast cancer to the gallbladder. Symptoms of cholecystitis should not be neglected in such patients, as they might indicate metastasis to the gallbladder.

Ciclooxigenase-2 nos carcinomas ductais de mama invasivos com componente ductal in situ e no epitélio adjacente

OBJETIVO: avaliar a presença da ciclooxigenase-2 (COX-2) nos carcinomas de mama ductal in situ (CDIS) e ductal invasivo (CDI), no estroma adjacente normal e no epitélio. A correlação entre os níveis de expressão com os graus nuclear e histológico, tamanho do tumor e idade da paciente foi também analisada. MÉTODOS: foram incluídos 47 espécimes cirúrgicos provenientes de mastectomias e quadrantectomias com CDI e CDIS concomitantes, estádios clínicos I e II. Foram utilizados anticorpos policlonais anti-COX-2 para determinar a expressão da enzima. As amostras foram categorizadas em escores de zero a três, de acordo com a intensidade e o número de células coradas. RESULTADOS: COX-2 foi positivamente expressa em CDI, CDIS e epitélio normal em 86,7, 84,4 e 73,3% dos casos, respectivamente. Quanto ao grau nuclear (GN), a expressão da COX-2 foi positiva em 80% dos casos de GN-I; 81,5 e 78,9% de GN-II; e 88,5 e 96,1% de GN-III no CDIS e CDI, respectivamente. A expressão da COX-2 ocorreu em 78,9% dos CDIS com comedonecrose e em 89,3% sem comedonecrose. Quanto ao grau histológico (GH) do CDI, a COX-2 foi positiva em 83,3% de GH-I; 89,9% de GH-II e 80% de GH-III. Em relação ao diâmetro tumoral, COX-2 esteve presente em 86,1% de CDI e 83,3% de CDIS em tumores com >2 cm de diâmetro e 11% em CDI; e CDIS em tumores ≤ 2 cm. A faixa etária ≥ 50 anos apresentou 90% de expressão para CDI e 86,7% para CDIS, e 92,5% de COX-2 para ambos CDI e CDIS em pacientes com idade <50 anos. CONCLUSÕES: nossos resultados demonstram alta correlação entre as expressões da COX-2 nos CDI, CDIS e epitélio normal, o que é consistente com a hipótese de que a superexpressão da COX-2 é um evento precoce na carcinogênese mamária. Não houve diferença quando da análise do grau histológico, presença de comedonecrose, grupo etário e tamanho do tumor.

Expression of MT-1 MMP, MMP2, MMP9 and TIMP2 mRNAs in Ductal Carcinoma in Situ and Invasive Ductal Carcinoma of the Breast

We investigated the expression of membrane type-1 (MT1)-MMP, MMP2, MMP9 and TIMP2 mRNAs and their roles in ductal carcinoma in situ (DCIS) and T1 and T2 invasive ductal carcinoma of the breast. We further compared these two types of carcinomas for differences in microvessel density, and expression of angiogenic factors and CD44std. MT1-MMP, MMP2, MMP9 and TIMP2 mRNA were expressed in both DCIS and invasive ductal carcinomas. Expression rates of MT1-MMP, MMP2, MMP9 and TIMP2 mRNAs were not statistically different between DCIS and invasive ductal carcinomas, nor did they differ statistically when grouped by tumor size, histologic grade or nuclear grade of invasive ductal carcinoma. Microvessel density and expression of VEGF and TGF-β1 were not statistically different between DCIS and invasive ductal carcinoma. CD44std expression was significantly increased in DCIS compared to invasive ductal carcinoma (p < 0.05) and it was also significantly increased in lower clinical stage, histologic grade and nuclear grade of invasive ductal carcinoma (p < 0.05). Axillary node metastasis was significantly correlated with MT1-MMP mRNA, VEGF and TGF-β1 expression (p < 0.05) and MT1-MMP mRNA was positively correlated with VEGF expression and TIMP2 mRNA (p < 0.05). In summary, patterns of MMP mRNA expression in DCIS and invasive ductal carcinoma suggest that the invasive potential of breast carcinoma is already achieved before morphologically overt invasive growth is observed. As MT1-MMP mRNA expression is significantly correlated with axillary nodal metastasis, it may be useful as a prognostic indicator of invasive ductal carcinoma. Considering the positive correlation of MT1-MMP mRNA and TIMP2mRNA expression, our finding supports a role for TIMP2 in tumor growth, as well as the utility of CD44std as a prognostic indicator of breast cancer.

Image analysis of irregularity of cluster shape in cytological diagnosis of breast tumors: Cluster analysis with 2D‐fractal dimension

To establish diagnostic criteria using comparison of cell cluster shapes, between benign and malignant tumors, breast tumors demonstrating weak cellular atypia in low grade invasive ductal carcinoma (IDC) were compared. Fine-needle aspiration (FNA) specimens of breast tumors were obtained from 37 patients. Among these, 16 were histologically diagnosed as IDC low-grade and the other 21 as benign fibroadenoma (FA). For evaluation, we examined 740 clusters from these 37 FNA specimens. Nine image morphometric parameters were studied, including the cluster area, circumference, maximal length, maximal breadth, ratio of length to breadth, cluster roundness, cluster size, and the edge and distribution image fractal dimensions for cluster analysis. We evaluated the irregularity in cell cluster shape using fractal dimension analysis, and determined the correlation to cluster size. The irregularity in the IDC cluster shape was higher than that in the FA cluster shape. However, six cases (28.5%) of 21 FA clusters showed high fractal dimensions similar to those for IDC. The clusters were classified by cluster analysis into three types: IDC clusters, FA with irregular cluster shape, and FA with no irregular clusters. The average cell cluster area of the FA with irregular shape was found to be about three times larger than that of IDC clusters. When the differential diagnosis between IDC and FA is difficult, it is important to focus on irregularities in the shape and on overall size of the cell clusters. For accurate diagnosis, the cell cluster shape is as important as the individual cellular atypia.

Invasive lobular carcinoma: to grade or not to grade

Grading of invasive ductal carcinoma of no special type using the Nottingham combined histologic grading system provides independent prognostic information. The prognostic utility of grading invasive lobular carcinomas, however, has not been fully elucidated. In addition, the relationship between grade in invasive lobular carcinomas and expression of predictive biomarkers is less certain. The purpose of this study was to correlate histologic grade in invasive lobular carcinoma with known prognostic and predictive markers. All primary resections for invasive mammary carcinomas diagnosed in Mount Sinai Hospital, Toronto, between the years 1996 and 2002 were reviewed (n=1053). Of these cases, 50 were pure invasive lobular carcinoma (incidence 4.7%). The median age at diagnosis was 64 years. These tumors were graded using the Nottingham combined histologic grading system and analyzed for estrogen receptor, progesterone receptor, HER2/neu and E-cadherin expression. Tumor grade was correlated with tumor size (P=0.03), and the American Joint Committee on Cancer nodal status (P=0.05). Assessment of the individual components of grade showed that the mitotic score was highly correlated with tumor size (P=0.02), lymph node positivity (P=0.02) and overall American Joint Committee on Cancer stage (P=0.01). Estrogen receptor and progesterone receptor were highly expressed irrespective of the grade of tumor. HER2/neu protein overexpression and E-cadherin protein expression was absent in all invasive lobular carcinomas studied. We conclude that pure invasive lobular carcinoma is uncommon and occurs predominantly in postmenopausal women. Increasing tumor grade is correlated with median tumor size and the American Joint Committee on Cancer nodal stage, but not correlated with the expression of estrogen receptor, progesterone receptor, E-cadherin or HER2/neu protein overexpression.

Evaluation of pathways for progression of heterogeneous breast tumors

To better understand the progression of heterogeneous breast cancers, four models of progession pathways have been evaluated. The models describe the progression through the grades of ductal carcinoma in situ (DCIS) 1, 2, and 3, and through the grades of invasive ductal carcinoma (IDC) 1, 2, and 3. The first three pathways, termed linear, nonlinear, and branched, describe DCIS as a progenitor of IDC, and grades of DCIS progressing into grades of IDC. The fourth pathway, termed parallel, describes DCIS and IDC as diverging from a common progenitor and progressing through grades in parallel. The best transition rates for the linear, nonlinear, and branched pathways were sought using a random search in combination with a directed search based on the Nelder–Mead simplex method. Parameter values for the parallel pathway were determined with heuristic graphs. Results of computer simulation were compared with clinically observed frequencies of grades of DCIS and grades of IDC that were reported to occur together in heterogeneous tumors. Each of the four pathways could simulate frequencies that resembled, to varying degrees, the clinical observations. The parallel pathway produced the best correspondence with clinical observations. These results quantify the traditional descriptions in which grades of DCIS are the progenitors of grades of IDC. The results also raise the alternative possibility that, in some tumors with both components, DCIS and IDC may have diverged from a common progenitor.