Grade Invasive Ductal Carcinoma Research Articles

Abstract P4-04-03: A microgenomic approach using LCM procured breast cancer cells to identify clinically relevant gene signatures that underlie favorable response of ILC to aromatase inhibitors

Abstract Based on CGH analyses, invasive lobular carcinoma (ILC) is closely related to low grade invasive ductal carcinoma (IDC), and is genetically unrelated to intermediate and high grade invasive carcinomas (Pathol Res Pract 2005; 201:713). Results from the BIG 1-98 trial, showed that patients with ILC and luminal B (intermediate/high grade) IDC treated with Letrozole, an aromatase inhibitor (AI), had a significant improvement in overall survival, whereas patients with luminal A (low grade) IDC did not (Cancer Res 2012; 72(24 Suppl):Abstract nr S1-1). Despite genetic evidence suggesting a close relationship between ILC and low grade IDC, and disparity between ILC and intermediate and high grade IDC, a clinically relevant gene set underlying a tumor's biologic responsiveness to AIs likely exists. We are deciphering a gene subset that would differ in expression between low grade IDC and ILC, and would be shared by ILC and luminal B IDC. Identification of such a gene set should lead to a more robust marker for identifying AI responsive tumors rather than relying solely on histomorphology and/or immunohistochemistry for the pathologic diagnosis of ILC, since the distinction between ILC and IDC is not always straight forward in pathology practice. Using 247 de-identified human breast carcinoma biopsies collected under standardized, stringent conditions, total RNA was extracted from carcinoma cells procured by laser capture microdissection to perform microarray analyses of approximately 22,000 genes to identify expression signatures associated with breast cancer characteristics. Of the 247 samples, 169 samples were used for the analysis and included 16 ILCs, 13 low grade IDCs, 55 intermediate grade IDCs and 85 high grade IDCs. Preliminary analyses identified 1267 genes that were differentially expressed (p<0.01) between cell samples from patients with IDC compared to those with ILC. Gene expression levels in breast cancer cells from patients with low grade IDC compared to those with ILC yielded 200 genes that were statistically differentially expressed (p<0.01). When comparing high grade IDC to ILC, 149 of these genes were not differentially expressed indicating that this gene set may serve as a panel of candidate genes for identifying cancers responsive to AIs. Pathway analysis software (Ingenuity®) identified that these genes were associated with key molecular and cellular functions, e.g., cell-to-cell signaling, cell morphology, cell death and survival, cellular assembly and organization and carbohydrate metabolism. qPCR analyses are used to validate and refine the gene subset distinguishing ILC from low grade IDC. Our approach is revealing subtle molecular features that discriminate these lesions based on clinical behavior. Supported in part by a grant from the Phi Beta Psi Charity Trust. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-04-03.

Abstract P5-16-05: Comparative analysis of the immunohistochemical profile of ductal carcinoma in situ to invasive ductal carcinoma

Abstract Introduction: The molecular subtypes have classified invasive breast carcinomas in diverse entities with different clinical behaviors, but the prevalence of these subtypes in ductal carcinoma in situ (DCIS) has not been evaluated in detail. The main objectives of this study were to compare proteomic expression profiles of DCIS and invasive ductal carcinoma (IDC) by immunohistochemistry (IHC), classify them according to the molecular subtypes and evaluate the relationship between the expression of the biological markers and the tumoral grade. Methods: We assessed the frequency of expression of ER, PR, HER2, proliferation markers (PCNA or Ki67), Bcl-2 and p53 in 107 DCIS and compared them with the expression of 682 IDC. They were classified according to molecular subtypes. We evaluated the relationship between the expression of these markers and the nuclear and histological grades of DCIS and IDC respectively. Results The expression of Bcl-2 and PR was significantly more frequent in the DCIS group (p = 0.0461 and p = 0.0001 respectively). The IDC showed significantly increased values for cell proliferation markers and mutated p53 (p < 0.0001 and p = 0.0062 respectively). The prevalence of ER and HER2 was similar in both groups (p = 0.0912 and p = 0.4686 respectively). Clinico-pathological parameters and biomarkers expression DCISIDCPN107682 Age (mean)52.75 years55.79 years0.0219Tumoral size (mean)1.62 cm2.23 cm<0.0001Grade 144 (41.12%)95 (13.92%)<0.0001Grade 229 (27.10%)277 (40.61%)0.0076Grade 334 (31.78%)310 (45.45%)0.0086ER positive82 (76.63%)467 (68.47%)0.0912PR positive70 (65.42%)373 (54.69%)0.0461HER2 positive19 (17.78%)101 (14.81%)0.4686Proliferation markers high65 (60.75%)206 (30.21%)<0.0001Bcl-2 positive84 (78.51%)405 (59.38%)0.0001p53 mutated33 (30.84%)307 (45.01%)0.0062 The luminal A subtype was more common in DCIS (p = 0.0003), whereas luminal B no-HER2 and triple negative were more prevalent in IDCs (p = 0.0195 and p = 0.0351 respectively). There were no differences in the frequency of luminal B-HER2 and HER2 positives (p = 0.3279 and p = 1.0000 respectively). Prevalence of molecular subtypes. DCISIDCPLuminal A46 (43%)172 (25.22%)0.0003Luminal B32 (29.91%)288 (42.23%)0.0195Luminal B-HER2+11 (10.28%)50 (7.33%)0.3279HER28 (7.47%)51 (7.48%)1.0000Triple Negative10 (9.34%)121 (17.74%)0.0351 In relation to nuclear grade of DCIS, no differences were detected with respect to the expression of ER, Bcl-2 and mutated p53 (p = 0.3691, p = 0.8136 and p = 0.3138 respectively). Proliferation markers increased significantly with increasing nuclear grade (p <0.0001). PR was mostly expressed in DCIS grade 1 and HER2 in grade 3, but this was not statistically significant (p = 0.0741 and p = 0.0851 respectively). With regard to histological grade of IDCs, there were a higher frequency of ER, PR, Bcl-2 in grades 1 (p <0.0001). G3 carcinomas showed increased expression of HER2, proliferation markers and mutated p53 (p <0.0001, p <0.0001 and p = 0.0110 respectively). Conclusions: The DCIS presents significant differences in tumor subtypes compared with IDC. The lower frequency of triple negative tumors and Luminal B-HER2-, lower expression of mutated p53 and lower degree of proliferation, suggest a less aggressive behavior of DCIS. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P5-16-05.

Abstract P4-01-13: Fusion imaging using opto-acoustics and ultrasound to differentiate breast mass histopathology

Abstract Introduction: Opto-acoustics (OA) is a true fusion imaging technology that combines dual energy laser optic functional imaging with real time gray scale ultrasound morphologic imaging. The laser optic component uses two wavelengths that have been chosen to optimally demonstrate oxygenated and deoxygenated hemoglobin. The color-coded laser optic images are co-registered and temporally interleaved with real time gray scale images. OA fusion imaging benefits from the combination of the high contrast resolution of dual wavelength laser optics and the higher spatial resolution and penetration of gray scale ultrasound. OA can demonstrate both tumor neovascularity and the relative degrees of oxygenation-deoxygenation within breast mass and within the surrounding tissues. Methodology: A series of 68 prospective patients with undiagnosed breast masses going to biopsy were evaluated before biopsy with a handheld device combining ultrasound co-registered with OA for the visualization of pre-defined internal and external features. The internal features included vascularity (V), blush (BU), and blood (BL) accumulation while external features included boundary zone vascularity and blush (BZ) and peri-tumoral radiating vessels (RV); each were scored on a 0-5 ordinal scale and were summed to get a total internal score, total external score, and a total score. A two-sided exact Jonckheere-Terpstra test was used to test the relationship between increasing scores and higher cancer grade. Results: The study population consisted of 29 fibroadenomas (FAs) as well as 10 invasive ductal carcinoma (IDC), grade 1, 11 IDC grade 2, and 18 IDC grade 3 cases. OA Total (two-sided p = 3×10−8) and OA Total Internal (two-sided p = 6×10−9) were highly significant but OA Total External was not statistically significant; OA Total and OA Total Internal Features were highly related to histology with lower scores for FAs increasing progressively to highest scores for IDC 3. Moreover, all three internal OA features were highly significant: (1) V (two-sided p = 10−7), BU (two-sided p = 3×10−8), and BL (two-sided p = 5×10−6). Cumulative contribution of each internal feature to the total internal score was also assessed for each histopathology subgroup to confirm independent contribution to the OA Total Internal Score. Pairwise correlations explained no more than 56% of the total variance. Conclusions: We have established the prognostic value of OA Total and OA Internal Features as well as the validation of the contributions of each OA Internal Feature. Using the combination of individual internal and external OA features not only helps distinguish benign from malignant masses, but helps stratify malignant lesions by histologic grade. These abilities could be useful both for diagnosis and for monitoring non-surgical treatment of malignant masses. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr P4-01-13.

Abstract ES03-1: Clinical 101: The pathology of breast cancer

Abstract Numerous histological types of breast cancer are recognized based on their discrete architectural patterns and associated different risk factors, clinical presentations, pathological features, patterns of spread, response to therapy and outcomes. Traditional prognostic factors include tumor size, lymph node status, tumor grade, and immunohistochemical profile of Estrogen and Progesterone Receptor (ER and PR) and Her2/neu gene amplification status. Histological tumor grade assesses degree of differentiation (tubule formation and nuclear pleomorphism) and proliferative activity (mitotic index). The three histological grades mirror aggressiveness and correlate with survival. Expression profiling reveals only two genomic grades, low and high. The ER positive, HER2 positive and triple negative tumors have correlative molecular subtypes on transcriptome analysis; two ER positive groups, luminal A and B where B tumors have a high proliferation compared with A, a HER2 enriched group, and basal-like group corresponding to triple negative tumors. A molecular apocrine group is characterized by ER negative, Androgen Receptor positive status with paradoxical expression of ER related genes and often HER2. A subset of molecular apocrine carcinomas have a morphological correlate. The traditional prognostic factors used to determine therapy neither capture the complexity of breast cancer nor provide tailored enough treatment options for individual patients. Gene expression profiles which predict disease recurrence pick out the proliferation related genes and a number of different prognostic signatures despite them having < 25% of shared genes between the signatures (e.g. OncotypeDX, PAM50, Mammaprint) identify the Luminal A low risk subgroup, which does not benefit from chemotherapy. These prognostic signatures have no value in ER negative disease where there is a high rate of resistance to chemotherapy regimens. Predictors of sensitivity or resistance to specific regimens of chemotherapy in all tumors and predictors of resistance to hormone therapy in ER positive tumors are needed. The low and high grade genomic signatures reflect two distinct models of tumor progression. The low grade neoplasia pathway is characterized by simple diploid karyotypes, and luminal A type invasive carcinomas in this group share genetic and transcriptomic features with precursor lesions, ADH, FEA. Special subtype tumors in this pathway include tubular, lobular, cribriform, mucinous carcinomas and some neuroendocrine carcinomas. The high grade pathway encompasses high grade DCIS, and high grade invasive ductal carcinoma and is characterized by aneuploidy and complex karyotypes. Although tumors can be lumped broadly into low and high grade, there is enormous diversity between tumors even within the molecular subgroups. For example the basal-like/triple negative group includes tumors with distinct morphology, clinical presentation, prognosis and molecular profiles which requires tailoring of therapies. Two rare low grade clinically indolent basal-like tumors include Secretory carcinomas which harbor a characteristic recurrent balanced chromosomal translocation which leads to a fusion transcript ETV6-NTRK3 and Adenoid cystic carcinomas which have a MYB-NFIB fusion gene. Citation Information: Cancer Res 2013;73(24 Suppl): Abstract nr ES03-1.

Nuclear loss of protein arginine N-methyltransferase 2 in breast carcinoma is associated with tumor grade and overexpression of cyclin D1 protein

Human protein arginine N-methyltransferase 2 (PRMT2, HRMT1L1) is a protein that belongs to the arginine methyltransferase family, and it has diverse roles in transcriptional regulation through different mechanisms depending on its binding partners. In this study, we provide evidences for the negative effect of PRMT2 on breast cancer cell proliferation in vitro and in vivo. Morever, cyclin D1, one of the key modulators of cell cycle, was found to be downregulated by PRMT2, and PRMT2 was further shown to suppress the estrogen receptor α-binding affinity to the activator protein-1 (AP-1) site in cyclin D1 promoter through indirect binding with AP-1 site, resulting in the inhibition of cyclin D1 promoter activity in MCF-7 cells. Furthermore, a positive correlation between the expression of PRMT2 and cyclin D1 was confirmed in the breast cancer tissues by using tissue microarray assay. In addition, PRMT2 was found to show a high absent percentage in breast caner cell nuclei and the nuclear loss ratio of PRMT2 was demonstrated to positively correlate with cyclin D1 expression and the increasing tumor grade of invasive ductal carcinoma. Those results offer an essential insight into the effect of PRMT2 on breast carcinogenesis, and PRMT2 nuclear loss might be an important biological marker for the diagnosis of breast cancer.

Ki-67 expression and significance of different molecular subtypes of breast invasive ductal carcinoma

To analyze Ki-67 expression and explore its significance in different molecular subtypes of breast invasive ductal carcinoma (IDC). The expressions of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor-2 (HER-2) and Ki-67 were detected in 126 cases of IDC by immunohistochemical staining. Then the molecular subtype of each case of IDC was determined.Statistical analysis was performed to determine the relationship between Ki-67 expression and the molecular subtypes with clinicopathological features of IDC. There was no statistically significant difference of Ki-67 expression in age and tumor size (P > 0.05).However, significant difference existed in histological grading and lymph node metastasis (P < 0.05). The expression level of Ki-67 was negatively correlated with ER expression (r = -0.273, P = 0.002) and PR expression (r = -0.242, P = 0.007) and positively with HER-2 expression (r = 0.245, P = 0.006) . A low expression of Ki-67 was in LumianlA subtype (17/17) and high expression in other molecular subtypes. Moreover, the rate of high expression (Ki-67 LI>50%) in each subtype progressively increased with the degree of molecular typing and Ki-67 expression in different molecular subtypes showed significant difference (P < 0.05). The expression level of Ki-67 is correlated with histological grading and molecular type of IDC. High expression of Ki-67 carries poor prognosis. Thus it is necessary to perform a variety of routine clinicopathological examinations, such as Ki-67, ER, PR and HER-2.

Thrombospondin-4 expression is activated during the stromal response to invasive breast cancer

The thromobospondins are a family of extracellular glycoproteins that are activated during tissue remodeling processes such as embryogenesis, wound healing and cancer. Thrombospondin-4 (THBS4) is known to have roles in cellular migration, adhesion and attachment, as well as proliferation in different contexts. Data to support a role in cancer biology is increasing, including for gastrointestinal and prostate tumours. Here, using a combination of immunohistochemistry, immunofluorescence and analysis of publicly available genomic and expression data, we present the first study describing the pattern of expression of THBS4 in normal breast and breast cancer. THBS4 was located to the basement membrane of large ducts and vessels in normal breast tissue, but was absent from epithelium and extracellular matrix. There was a significant induction in expression in cancer-associated stroma relative to normal stroma (P = 0.0033), neoplastic epithelium (P < 0.0001) and normal epithelium (P < 0.0001). There was no difference in stromal expression of THBS4 between invasive ductal carcinomas (IDC) and invasive lobular carcinomas (ILC). The THBS4 mRNA levels were variable yet were generally highest in tumours typically rich in stromal content (ILC, ER positive low grade IDC; luminal A and normal-like subtypes). Genomic alterations of the THBS4 gene (somatic mutations and gene copy number) are rare suggesting this dramatic activation in expression is most likely dynamically regulated through the interaction between invading tumour cells and stromal fibroblasts in the local microenvironment. In summary, THBS4 expression in breast cancer-associated extracellular matrix contributes to the activated stromal response exhibited during tumour progression and this may facilitate invasion of tumour cells.

A new method for apparent diffusion coefficient measurement using sequential 18F-FDG PET and MRI: correlation with histological grade of invasive ductal carcinoma of the breast

The aim of this study was to measure the apparent diffusion coefficient (ADC) value at the region with the highest FDG uptake using sequential (18)F-FDG PET and MRI, and to correlate it with the histological grade of invasive ductal carcinoma (IDC) of the breast. A retrospective study was conducted on 75 untreated patients with IDC. First, a PET/CT scan and subsequent breast MRI were done and the SUVmax of the each breast tumor was recorded. Then, a PET image and ADC map were co-registered. On the axial slice containing the pixel with SUVmax, we drew multiple circular ROIs within the tumor and measured the mean ADC value of each ROI. The average (ADC-mean) and minimum (ADC-min) of the mean ADC values for all ROIs within the tumor were calculated, respectively. Then, a circular ROI was placed at the corresponding location to the pixel with the highest SUV and the mean ADC value of the ROI was denoted as ADC-PET. We compared the averages of the ADC parameters and assessed the correlations among SUVmax and ADC parameters. ROC curve and logistic regression analyses were performed to assess the utility of ADC and SUVmax for detecting histological grade 3. ADC-min was significantly lower than the ADC-mean or ADC-PET. All of the ADC parameters showed a negative correlation with SUVmax. The area under the ROC curve for identifying histological grade 3 using ADC-PET, ADC-min, ADC-mean and SUVmax was 0.684, 0.660, 0.633 and 0.639, respectively. By multivariate analysis, ADC-PET was a significant, independent predictor of histological grade 3 (p = 0.004). We estimated the ADC value at the breast tumor region with the highest FDG uptake using sequential (18)F-FDG PET and MRI. This new ADC parameter distinguished high-grade IDC, supporting the feasibility of the combined PET-MRI system in patients with breast cancer.

Expression of fatty acid synthase and its association with HER2 in invasive ductal carcinoma of breast

To investigate the expression of fatty acid synthase (FAS) in adenosis, atypical ductal epithelial hyperplasia, ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) of breast, and the correlation of FAS expression with HER2 gene amplification in IDC. Immunohistochemical EnVision method staining for FAS was performed in 100 cases of breast lesions and 10 normal breast tissues. HER2 gene amplification was detected with FISH in 60 cases of IDC. The cohort included 10 cases of adenosis, 10 atypical ductal epithelial hyperplasia, 20 DCIS (8 high-grade, 9 intermediated-grade and 3 low-grade), and 60 cases of IDC (5 grade 1, 40 grade 2 and 15 grade 3). FAS expression was negative in all 10 normal breast tissues; in the 10 cases of adenosis, strongly positive FAS expression was detected in one case, positive in 2, weakly positive in 4, and negative in 3; in the 10 cases of atypical ductal epithelial hyperplasia, FAS immunohistochemistry showed that 1 was strongly positive, 4 positive, 4 weakly positive, and 1 negative; in the 20 cases of DCIS, FAS immunostaining showed that 12 were strongly positive, 5 positive, 1 weakly positive, and 2 negative; FAS expression showed a clear increasing trend from normal breast tissue, atypical ductal epithelial hyperplasia to DCIS (χ(2) = 42.02, P < 0.01). Likewise, the increasing trend was also demonstrated from adenosis to DCIS (χ(2) = 34.69, P < 0.01). There was also a positive correlation between FAS expression and extent of lesion among normal breast tissue, adenosis, atypical ductal epithelial hyperplasia and DCIS (χ(2) = 86.02, P < 0.01; r = 0.568, P < 0.01). FAS expression was not correlated with the grade of DCIS (χ(2) = 9.12, P = 0.16). In the five cases of grade 1 IDC, FAS immunostaining showed that 4 cases were strongly positive and 1 positive; in the 40 cases of grade 2 IDC, FAS immunostaining showed that 27 strongly positive, 12 positive, and 1 negative; in the 15 cases of grade 3 IDC, FAS immunostaining showed that 6 were strongly positive, 5 positive, 3 weakly positive, and 1 negative; FAS expression was stronger and more extensive in DCIS, IDC grades 1 and 2 than that in other groups. However, FAS expression was weaker in the IDC grade 3 (χ(2) = 11.26, P = 0.01). The positive expression rate of FAS in IDC was generally higher than that in benign breast lesions (χ(2) = 47.19, P < 0.01). In the 60 cases of IDC, FISH showed HER2 gene amplification in 22 cases, but not in the remaining 38 cases. FAS expression in IDC was highly correlated with HER2 gene amplification (r = 0.44, P < 0.01). The expression of FAS had significant correlation with status of ER and PR and tumor size (P < 0.05). There was no significant correlation with age, immunohistochemical HER2 expression, lymph node metastasis and clinical stage (P > 0.05). FAS may be closely related to the carcinogenesis of breast IDC. FAS expression is closely associated with HER2 gene amplification in IDC.

Is there different correlation with prognostic factors between “non-mass” and “mass” type invasive ductal breast cancers?

PurposeTo investigate the association between non-mass type breast cancer and common clinical–pathological prognostic factors, compared with mass type breast cancer. Materials and methodsAfter institutional review board approval, retrospective blind review of contrast-enhanced breast MRI was carried out for 88 histologically proven breast invasive ductal carcinoma (IDC) patients, presenting from January 2008 to December 2011. Two radiologists assessed the images of each lesion for the morphologic enhancement type [mass enhancement or non-mass-like enhancement (NMLE)] and the distribution/internal enhancement of NMLE. Two pathologists evaluated the histological grade of IDC, presence or absence of ductal carcinoma in situ (DCIS), lymph node status, presence or absence of vascular invasion, and expression status of estrogen receptor (ER)/progesterone receptor (PR)/HER-2/p53 tumor suppressor gene (p53)/Ki-67. Inter-observer agreement was assessed with kappa test. Chi-square test and Spearman rank correlation were performed to explore the associations of morphologic enhancement type with the age, lesion size and the above pathological prognostic factors ResultsInter-observer agreement was excellent, with kappa>0.75. Morphologic enhancement type was significantly correlated with age (P=0.02), with NMLE more commonly seen in women less than 50 y/o. The size of NMLE was larger than that of mass and, with the increase of lesion size, proportion of NMLE among the cases increased (P=0.001). NMLE was also significantly correlated with low histologic grade of IDC (P=0.003) and presence of DCIS (P<0.001). There was no significant correlation between morphologic enhancement type and lymph node status, vascular invasion, ER/PR/HER-2/p53/Ki-67 status. The histological grade was higher in clumped enhancement than non-clumped (P=0.011). There was no correlation between enhancement distribution and prognostic factors ConclusionsNon-mass type breast cancer may not necessarily have worse prognosis than the mass type, due to lower histological grade and closely related to DCIS component, although it may has larger tumor size. Clumped enhancement may have worse prognosis than non-clumped enhancement.

Diagnosis Error: Carcinoma or Primary Breast Lymphoma? A Case Report and Literature Review

Introduction: Breast involvement by non-Hodgkin lymphoma is particularly rare in women. We describe the case of a patient with a rapidly growing, nodule in the right breast. On ultrasonography, the nodule was suspicious for breast carcinoma. Case presentation: A breast biopsy from a 73-year-old Moroccan women answered invasive ductal carcinoma grade 3 from Elston and Ellis. Patey was performed. Microscopic examination showed lymphoid proliferation confirmed on immunohistochemical analysis. Our patient was treated with chemotherapy. Conclusions: The rarity of breast lymphomas, and the problems related to the diagnosis and therapeutic choices with these tumors require molecular techniques in association with classical histological diagnosis.

Relationship between ultrasound appearance and histologic grades and expression of Ki-67 in breast cancer

Objective To study the relationship between ultrasound appearance and histologic grades and expression of Ki-67 in breast cancer.Methods The ultrasound appearances of 104 breast carcinomas confirmed by pathology were analyzed.Immunohistochemical method was used to test the expression of Ki-67 in all the breast carcinomas analyzed the relationship between ultrasound appearance and histologic grades and expression of Ki-67 in hreast cancer.Results ( 1 ) The expression rate of Ki-67 were 73.1％ ( 76/104 ).( 2 ) Among the 82 cases of invasive ductal carcinoma( IDC ),appearance rate of diameter ≥ 2 cm and Ⅱ ～ Ⅲ grade flow was higher in Ⅲ grade IDC than in Ⅱ or Ⅰ grade IDC.( 3 )Appearance rate of diameter ≥2 cm,microcalcification and axillary lymph node metastasis were higher in Ki-67-positive tumors than in Ki-67-negative tumors ( P ＜ 0.05 ).( 4 ) Expression rate of Ki-67 was higher in Ⅲ grade IDC than in Ⅱ or Ⅰ grade IDC( P＜ 0.05 ).Conclusion To some extent,ultrasound appearance can reflect the histologic grade and expression of Ki-67 in breast cancer. Key words: Breast cancer; Ultrasound appearance; Histologic grades; Ki-67

Molecular Detection of Epstein-Barr virus in Breast Cancer

Background: The association of Epstein Barr virus (EBV) with breast carcinomas (BCs) is still in controversy. The aim of this study was to clarify the association of EBV & BC in Egyptian females and to assess its role as potential contributor to the development and behavioral alteration of BC. Subjects & methods: EBV-DNA was detected using PCR on breast tissue from 40 female patients with primary invasive BC; ductal (n=32) and lobular (n=8) and 20 age matched females undergoing reduction mammoplasties as control. Results: EBV-DNA was detected in 8/40 (20%) BC specimens. On the other hand all control specimens were negative As regards prognostic factors, no association was observed between EBV-DNA and patients' age, menopausal status and steroid receptor expression. However, significant associations were detected between the presence of EBV-DNA and other poor prognostic factors. All of the EBV-DNA positive BC were significantly associated with positive nodal status, where 7/8 cases showed more than three tumor-positive LN involvement. In spite of the small number of invasive lobular carcinoma included in this study there was a significant correlation between this histological type of poor prognosis and EBV-DNA detection rate where 4/8 (50%) of them were positive for EBV-DNA compared to 4/32 (12.5%) detection rate in invasive ductal carcinoma. A significant correlation was found between EBV-DNA detection rate in BC and high tumor grade of invasive ductal carcinoma; (100%, 1/1) association with grade III versus (9.67%; 3/ 31) with grade II. Conclusions: our results demonstrated the presence of the EBV genome in a considerable subset of BC in Egyptian patients. The virus was more frequently associated with bad prognostic factors. This indicates that EBV may play a role in the development and behavioral alteration of some aggressive BC.

Molecular Detection of Epstein-Barr virus in Breast Cancer

Background: The association of Epstein Barr virus (EBV) with breast carcinomas (BCs) is still in controversy. The aim of this study was to clarify the association of EBV & BC in Egyptian females and to assess its role as potential contributor to the development and behavioral alteration of BC. Subjects & methods: EBV-DNA was detected using PCR on breast tissue from 40 female patients with primary invasive BC; ductal (n=32) and lobular (n=8) and 20 age matched females undergoing reduction mammoplasties as control. Results: EBV-DNA was detected in 8/40 (20%) BC specimens. On the other hand all control specimens were negative As regards prognostic factors, no association was observed between EBV-DNA and patients' age, menopausal status and steroid receptor expression. However, significant associations were detected between the presence of EBV-DNA and other poor prognostic factors. All of the EBV-DNA positive BC were significantly associated with positive nodal status, where 7/8 cases showed more than three tumor-positive LN involvement. In spite of the small number of invasive lobular carcinoma included in this study there was a significant correlation between this histological type of poor prognosis and EBV-DNA detection rate where 4/8 (50%) of them were positive for EBV-DNA compared to 4/32 (12.5%) detection rate in invasive ductal carcinoma. A significant correlation was found between EBV-DNA detection rate in BC and high tumor grade of invasive ductal carcinoma; (100%, 1/1) association with grade III versus (9.67%; 3/ 31) with grade II. Conclusions: our results demonstrated the presence of the EBV genome in a considerable subset of BC in Egyptian patients. The virus was more frequently associated with bad prognostic factors. This indicates that EBV may play a role in the development and behavioral alteration of some aggressive BC.

Myoepithelial expression of Fas and strong nuclear expression of FasL in epithelial cells: A marker for risk stratification of breast cancer

The clinical significance of Fas and FasL in hormone-sensitive carcinomas has been reported. Our objective was to evaluate the expression of apoptosis-regulating genes Fas and FasL in Indian breast cancer and fibroadenoma patients in relation to hormone receptor status. Retrospective. Paraffin-embedded tissue samples from 63 untreated female patients with invasive ductal carcinoma (IDC) and 32 female patients with fibroadenoma were studied. Expression of Fas and FasL was evaluated using immunohistochemical staining method. Fisher's exact test and nonparametric correlation test (Spearman rank correlation test) were performed. Fas was detected in 97% of the fibroadenomas and there was a slight decrease in levels of expression with histological grades of IDC. The expression of FasL was detected in 75% fibroadenomas and its expression increased in IDC. There was no correlation between Fas, FasL expression and hormone receptor status. Strong expression of Fas in myoepithelial cells was noted in 12 out of 32 fibroadenoma cases. Expression of Fas and FasL alone is unlikely to be important as a predictive factor as they express in both normal and malignant breast epithelium. But strong expression of Fas in myoepithelial cells along with strong nuclear expression of FasL in epithelial cells of fibroadenoma could be useful as an early predictive factor for onset of malignancy.

Expression of the Androgen Receptor and its Correlation with Molecular Subtypes in 980 Chinese Breast Cancer Patients

BackgroundRecent studies have shown that androgen displays an inhibitory effect on breast cancer cell lines that express androgen receptor (AR) but not estrogen receptor (ER) and progesterone receptor (PR). We have previously reported that approximately 1/3 of ER negative high grade invasive ductal carcinomas express AR. Thus, AR can serve as a potential therapeutic target for this group of patients.AimHere we investigated AR expression patterns in 980 consecutive breast carcinomas.ResultsWe found that (1) AR was expressed more frequently (77%) than ER (61%) and PR (60%) in breast carcinomas; (2) AR expression was associated with ER and PR expression (P < 0.0001), small tumor size (P = 0.0324) and lower Ki-67 expression (P = 0.0013); (3) AR expression was found in 65% of ER negative tumors; (4) AR expression was associated with PR and Ki-67 in ER negative tumors, but not in ER positive tumors; (5) AR expression was higher in ER positive subtypes (Luminal A, Luminal B and Luminal HER2 subtypes, 80%–86%) and lower in ER negative subtypes [HER2, triple negative (TN), and TN EFGR positive subtypes; 52%–66%], with over 50% of TN tumors expressing AR.ConclusionMore breast carcinomas express AR than ER and PR, including significant numbers of ER negative and TN tumors, for which AR could serve as a potential therapeutic target.

Reduced expression of activin receptor-like kinase 7 in breast cancer is associated with tumor progression

To explore the clinical implication of activin receptor-like kinase 7 (ALK7) expression in breast cancer, we evaluated its protein level in six kinds of human breast tissue samples, including adjacent normal tissues, adenosis, breast fibroadenoma, ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC), and lymph node metastases (LNM). Immunohistochemical analyses showed that ALK7 was more frequently and much more intensely expressed in adjacent normal tissues, adenosis, and fibroadenoma tissues than in malignant tissues (DCIS, IDC, and LNM). Furthermore, the ALK7 expression in primary tumors and the corresponding LNM was evaluated in parallel samples from 60 patients with IDC. Results showed that the ALK7 expression status in primary tumors and LNM was concordant in 53 patients (88%), suggesting that ALK7 expression was retained in LNM. Moreover, our results suggested that ALK7 expression inversely correlated with the tumor grade (P=0.009) and clinical stage (P=0.004) in IDC significantly. Finally, the effect of activin-ALK7 pathway on the breast cancer cell growth was elucidated, and results revealed that overexpression of ALK7 could restore the inhibitory effect of activin B on the growth of ALK7-negative breast cancer cell line, ZR-75-30. These findings provide the evidence that the reduction or lack of ALK7 expression may account for the loss of its ligand sensitivity of breast cancer cells, thereby leading to breast tumor progression.

Loss of interferon regulatory factor 5 (IRF5) expression in human ductal carcinoma correlates with disease stage and contributes to metastasis

IntroductionNew signaling pathways of the interleukin (IL) family, interferons (IFN) and interferon regulatory factors (IRF) have recently been found within tumor microenvironments and in metastatic sites. Some of these cytokines stimulate while others inhibit breast cancer proliferation and/or invasion. IRFs, a family of nine mammalian transcription factors, have multiple biologic functions that when dysregulated may contribute to tumorigenesis; most well-known are their roles in regulating/initiating host immunity. Some IRF family members have been implicated in tumorigenesis yet little is still known of their expression in primary human tumors or their role(s) in disease development/progression. IRF5 is one of the newer family members to be studied and has been shown to be a critical mediator of host immunity and the cellular response to DNA damage. Here, we examined the expression of IRF5 in primary breast tissue and determined how loss of expression may contribute to breast cancer development and/or progression.MethodsFormalin-fixed paraffin-embedded archival breast tissue specimens from patients with atypical ductal hyperplasia (ADH), ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) were examined for their expression of IRF1 and IRF5. Knockdown or overexpression of IRF5 in MCF-10A, MCF-7 and MDA-MB-231 mammary epithelial cell lines was used to examine the role of IRF5 in growth inhibition, invasion and tumorigenesis.ResultsAnalysis of IRF expression in human breast tissues revealed the unique down-regulation of IRF5 in patients with different grades of DCIS and IDC as compared to IRF1; loss of IRF5 preceded that of IRF1 and correlated with increased invasiveness. Overexpression of IRF5 in breast cancer cells inhibited in vitro and in vivo cell growth and sensitized them to DNA damage. Complementary experiments with IRF5 siRNAs made normal mammary epithelial cells resistant to DNA damage. By 3-D culture, IRF5 overexpression reverted MDA-MB-231 to normal acini-like structures; cells overexpressing IRF5 had decreased CXCR4 expression and were insensitive to SDF-1/CXCL12-induced migration. These findings were confirmed by CXCR4 promoter reporter assays.ConclusionsIRF5 is an important tumor suppressor that regulates multiple cellular processes involved in the conversion of normal mammary epithelial cells to tumor epithelial cells with metastatic potential.

Immunophenotyping analysis in invasive micropapillary carcinoma of the breast: Role of CD24 and CD44 isoforms expression

We analyzed immunohistochemically the expression of CD24 and spliced variants of CD44v5 and v9 in invasive micropapillary carcinoma (IMPC) of the breast that is a rather aggressive tumor characterized by alteration of cells adhesion molecules, early lymph node metastases and poor prognosis. We analyzed 31 high-grade IMPCs and compared their expression to 22 high grade (G3) invasive ductal carcinomas of the breast (IDCs). We found a higher expression of CD24 in high-grade IMPCs with a peculiar inverted apical localization, compared to IDCs, showing a strong cytoplasmic staining; normal breast tissue resulted completely negative. IMPCs showed reduced expression of CD44v5 and CD44v9 compared with IDCs, but without a statistical significant difference. This study demonstrated that IMPC represents a distinct entity of breast carcinoma with high expression of CD24 with a typical inverted apical membrane pattern and reduction of CD44 isoforms v5 and v9, compared to IDCs. These features could explain the high lymph-vascular invasion propensity and higher metastatic capability of these tumors and could be a useful tool for a future targeted therapy.

Increased expression of fibroblastic growth factor receptor 2 is correlated with poor prognosis in patients with breast cancer

Although there is growing evidence supporting the hypothesis that fibroblast growth factor receptor 2 (FGFR2) is one of the few candidate genes linked with breast cancer susceptibility, the precise role of FGFR2 protein expression in breast cancer is still unknown. Our study examines FGFR2 protein expression in breast cancer and determines its associations with clinicopathological features and survival. Specimens from 125 invasive ductal carcinoma grade 2 (IDC2) breast cancer patients were investigated by immunohistochemistry for FGFR2 protein expression. Associations between the expression of FGFR2 and various clinicopathological features as well as survival status were studied. Cytoplasmic and nuclear FGFR2 were expressed in 64.8% and 56.8% of breast cancer patients, respectively. Cytoplasmic FGFR2 expression was significantly associated with tumor size and TNM stage. Furthermore, patients with high expression levels of cytoplasmic and nuclear FGFR2 showed much lower overall survival (OS) and disease-free survival (DFS) rates than those patients with low FGFR2 expression. Cytoplasmic FGFR2 expression and lymph node metastasis were independent prognostic factors for both DFS and OS by multivariate analysis. High FGFR2 expression is correlated with poor OS and DFS in breast cancer patients. It could be a biomarker for poor prognosis.