Cases Of Normal Breast Tissue Research Articles

Association of chemotactic factor receptor 5 gene with breast cancer

We designed a 2-stage study to investigate chemotactic factor receptor 5 (CCR5) gene expression in breast cancer tissues and axillary lymph nodes and analyze the association between the CCR5-Î"32 gene polymorphism and the clinical features and prognosis of breast cancer patients. The first stage examined 72 cases of invasive ductal carcinoma and axillary lymph node tissue, 50 cases of breast fibroadenoma tissue, and 40 cases of normal breast tissue. The tissues specimens were embedded in paraffin, and CCR5 expression was detected using immunohistochemical methods. C-erbB-2, p53, Ki-67, estrogen receptor, and progesterone receptor expression were also detected in the breast cancer tissues. The second stage examined 35 cases of surgically removed tissue. Relative expression levels of CCR5 messenger RNA (mRNA) in primary foci, axillary lymph node, and cancer-adjacent tissues of the breast cancer and breast fibroadenoma samples were detected using real-time quantitative reverse transcription-polymerase chain reaction assay. We found that 1) CCR5 mRNA relative expression levels in breast cancer tissue were significantly higher than those in adjacent normal tissue (P < 0.01) and benign tumors (P < 0.05). The relative CCR5 mRNA relative expression level between phase II and phase III breast cancer tissues was statistically significant (P < 0.05). The CCR5 mRNA relative expression level between adjacent normal tissues and fibroadenoma tissues was not significantly different (P > 0.05). 2) Relative CCR5 mRNA expression level was significantly higher in metastatic lymph node tissues than that in non-metastatic lymph nodes (P < 0.05), and 3) CCR5 expression in breast cancer tissue was positively correlated with axillary lymph node metastasis (chi-square = 4.982, P = 0.026, r = 0.305). CCR5 expression was mildly and positively correlated with the oncogene C-erbB-2 (P < 0.05, r = 0.291). 4) CCR5 expression in breast cancer tissue was not correlated with age, menopause, maximum tumor size, tumor phase, p53, Ki-67, estrogen receptor, progesterone receptor, or other clinical features (P > 0.05). We concluded that CCR5 expression significantly increases in breast cancer tissues and metastatic lymph nodes. CCR5 plays a role in breast cancer development and axillary lymph node metastasis. It can be used indirectly as an indicator of axillary lymph node metastasis and prognosis.

Survivin expression in breast lobular carcinoma: Correlations with normal breast tissue and clinicomorphological parameters

The antiapoptotic protein survivin is rarely expressed in normal adult differentiated tissues, but it is often detected in their malignant counterparts. Immunohistochemically, we evaluated survivin expression in 19 cases of normal breast tissue and 64 cases of lobular breast carcinoma. The intensity of staining, percentage of labeled cells and subcellular location of survivin were assessed. We analyzed the quantitative differences of survivin expression between normal breast tissue and carcinomas. We also correlated survivin expression pattern in carcinomas with clinicomorphological parameters such as age of patients, grade, stage and size of primary tumor, lymph node metastasis, vascular invasion as well as estrogen and progesterone status. Survivin was detected in 10/19 cases of normal breast tissue (52.6%) and in 55/64 cases of lobular breast carcinoma (86%). The statistical analysis confirmed significant correlations between the assessed parameters in normal breast and lobular carcinoma. Furthermore, the expression of estrogen correlated significantly with the subcellular localization and intensity of survivin in carcinoma. However, no significant correlation was shown with regard to other clinicomorphological parameters. Our results suggest that survivin may be a valuable diagnostic marker, as well as a new independent prognostic parameter, in lobular breast carcinoma. Finally, our data support the hypothesis that lobular and ductal breast carcinomas seem to be different clinicomorphological entities.

Clinical Implications of p57KIP2Expression in Breast Cancer

To study the relationship between expression of p57KIP2 and prognosis and other clinicopathological parameters in invasive breast cancers. We assessed the expression of p57KIP2 in 89 cases of invasive breast cancer and 20 cases of normal breast tissue by immunohistochemical methods and analyzed the results with SPSS software (ver. 16.0). The positive expression rates of p57KIP2 protein in the invasive breast cancers and surrounding normal tissue were 30.3% (27/89) and 65% (13/20), respectively. Cases with no p57KIP2 expression exhibited a significantly higher post-operative distant metastasis rate than those with p57KIP2 expression (37.9% vs. 14.8%; P = 0.01). DFS analysis showed that p57KIP2-/C-erbB-2μ tumors also exhibited a significantly higher post-operative distant metastasis rate than the other groups (66.7% vs. 29.2%; P = 0.007), as did p57KIP2-/p53μ tumors (64.3% vs. 22.7%; P = 0.001). Survival analysis revealed that p57KIP2 was associated with breast cancer-specific survival overall (P = 0.045, log-rank test). Subgroup analysis demonstrated that individuals with p57KIP2-/C-erbB-2μ tumors experienced significantly worse post-operative survival than those with p57KIP2- /C-erbB-2- or other tumors (P = 0.006, log-rank test). p57KIP2-/p53μ tumors were associated with significantly worse post-operative survival than p57KIP2-/p53- or other tumors (P = 0.001, log-rank test). Cox regression analysis showed that p57KIP2 was a non-independent prognostic factor for breast cancer (P = 0.303). p57KIP2 is expressed at low levels in invasive breast cancer and is associated with better overall survival rate and disease-free survival in breast cancer patients, but it was a non-independent prognostic factor for breast cancer. Thus, the connection between p57KIP2/p53 and p57KIP2/C-erbB-2 may provide biomarkers for breast cancer.

Asymmetries: Comparison Between Full Field Digital Mammography and Digital Breast Tomosyntesis

INTRODUCTION: Breast Tomosynthesis (DBT) is a procedure developed from digital mammography unit to enable the acquisition of a three-dimensional (3D) volume of thinsection data. The slice examination of breasts can reduce or even eliminate tissue overlap. The purpose of this study was to evaluate if breast tomosynthesis can help to differentiate the assymetries from the overlapping tissues. MATERIALS AND METHODS: A total of 48 cases of asymmetries were referred to perform stereotactic guided core needle biopsy or vacuum assisted biopsy. Each case was analyzed with full field digital mammography(FFDM) and with digital breast tomosynthesis (DBT) previously to the procedure. The study included 47 women (age range 34-78 years, mean age 44, 3 years;) and all the lesions were sampled. The histopathologic results were compared with those obtained with FFDM and DBT. RESULTS: of the 48 cases of asymmetries, 19 (39, 58%) did not change with DBT and 27 (60, 41%) attenuated or disappeared. Among the 19 lesions, the stereotactic guided biopsy diagnosed 8 cases of breast cancer (4 invasive ductal carcinoma, 3 invasive lobular carcinoma, 1 ductal carcinoma in situ), 1 of lobular neoplasia, 1 atypical ductal hyperplasia, 6 of fibroadenoma and 3 cases of normal breast tissue. There were no cases of breast cancer or atypical hyperplasia in those images that attenuated or disappeared with DBT. In this study the biopsy could be avoided in 60, 41% of the cases of asymmetries with the performance of the DBT. DISCUSSION: These results indicate that DBT can be useful in the differentiation between the asymmetries and the overlapping tissues, allowing a decrease in the number of unnecessary biopsies in these cases.

Overexpression of Hedgehog signaling molecules and its involvement in triple-negative breast cancer

The purpose of this study was to investigate the activation of Hedgehog (Hh) signaling molecules and its involvement in triple-negative breast cancer (TNBC). A total of 123 cases of paraffin blocks, including 83cases of primary breast carcinoma, 30 cases of mammary hyperplasia and 10cases of normal breast tissue, were immunohistochemically analyzed for Sonic Hedgehog (SHH), Patched-1 (PTCH1), Smoothened (SMO) and glioma-associated oncogene homoglog1 (GLI1) expression. The expression of SMO and GLI1 in TNBC was significantly increased in comparison to non-triple-negative breast cancer (nTNBC). GLI1 expression manifested an inverse association with the estrogen receptor. The levels of GLI1 expression were increased in lymph node-positive cases. The expression of SHH and SMO was increased in high histological grades. Furthermore, the expression of SMO and GLI1 was correlated with superior tumor stage. The expression of SHH, SMO and GLI1 was significantly increased in breast cancer and mammary hyperplasia. PTCH1 expression was significantly decreased in breast cancer compared to mammary hyperplasia and normal breast tissue. For the first time, clinical evidence has been provided in support of significant roles of Hh signaling in TNBC. Hh signaling is involved in breast ductal changes and malignant transformation. Measures to inhibit Hh activity may improve the prognosis of TNBC patients.

Study on β-catenin gene expression and mutation in breast carcinoma

Objective To discuss the expression and mutation of β-catenin gene in breast carcinoma.Methods 119 breast carcinomas, 72 adenosis of breast tissues and 40 normal breast tissues were studied.Using immunohistochemistry studies to detect the expressions of β-catenin, then analyzing their relationship with the clinicopathological features of breast carcinomas. Gene mutation of β-catenin (exon 3) in 44 cases of breast carcinoma were analyzed by polymerase chain reaction (PCR) and then by direct sequencing.Results In 40 cases of normal breast tissues, epithelial cells showed equally strong membranous of β-catenin protein at the cell-cell boundaries. The abnormal rate of β-catenin in breast carcinomas (78.15 %) was much higher than that of adenosis of breast tissues (44.44 %), the significant correlation was found (P＜0.01). The abnormal expression of β-catenin was associated with lymph node metastasis (P=0.031). Mutation of β-catenin gene was not detected in 44 cases of breast carcinoma, while the abnormal rate of β-catenin in 44 cases of breast carcinoma was 77.30 %. Conclusion β-catenin abnormal expression could be considered as an useful marker for determining metastasis and prognosis of human breast carcinoma and for guiding treatment. β-catenin abnormal expression is not caused by β-catenin gene mutation, maybe by other mechanisms. It needs further study. Key words: Breast neoplasms; β-catenin; Mutation

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.

Expression of P53 and MDM2protein in breast tumor

Objective To investigate the expression of anti-oncogene P53 and cancergeneMDM2 proteins and their significance inthe pathogenesis and progress of Breast tumor. Methods To detect the expression ofP53 andMDM2 in 60 cases of Breast tumor and 20 cases of normal Breast tissue with immunoh is to chemical method. Results The positive rate of P53 in the tissues of the 60 cases of breast tumor is much higher than it in the 20 cases of normal breast tissues. There is statistical significance in the disparity (P ＜ 0. 01). Mdm2, P53protein expression and tumor size was no difference (P ＞ 0. 05) , clinical stage, axillary lymph node metastasis was significant difference(P ＜ 0. 01). Conclusion Both P53 andMDM2 participate in the pathogenesis and progress of breast tumor and they can be considered as the biological index to evaluate breast tumor. Key words: P53; MDM2; Breast tunor immunohistochemistry

Methylation and expression of gene p16INK4a and RB in breast carcinoma

(1) To investigate the promoter methylation status of gene p16(INK4a) and gene RB in breast carcinoma and the adjacent non-neoplastic hyperplastic epithelial tissue. (2) To study the correlation of p16(INK4a) gene expression at protein level with the abnormal gene methylation, the clinical manifestation and the pathological parameters. Methylation status of promoters of p16(INK4a) gene and RB gene was detected by using methylation specific PCR in 46 cases of breast cancer, 22 cases of the adjacent non-neoplastic hyperplastic epithelium tissue and 7 cases of normal breast tissue. In addition, the p16(INK4a) gene protein expression level was also detected using immunohistochemical technique(SP method) in 46 cases of breast cancer and 22 cases of the adjacent hyperplastic epithelial tissue. The methylation rate of p16(INK4a) gene was 23.9% (11/46) in breast cancer, 18.2% (4/22) in the adjacent non-neoplastic hyperplastic epithelial tissue and 1/7 in normal breast tissue, respectively. The methylation rate of RB gene was relatively low, which was 10.8% (5/46), 9.1% (2/22) and 0(0/7) in the above 3 groups, respectively. Methylation rate of p16(INK4a) gene and RB gene was not significantly different among the breast cancer, the adjacent non-neoplastic hyperplastic tissue and the normal tissues (P > 0.05). However, the methylation status of p16(INK4a) gene was closely correlated with its protein expression level and the negative ER expression result of the breast cancer (P < 0.05), but not correlated with the size of the cancer, differentiation status, lymph node metastasis, and age. The methylation status of RB gene was correlated with lymph node metastasis, but not with the size, the differentiation status, ER expression of the breast cancer and the age of the patients. The abnormal methylation of p16(INK4a) gene may not play a significant role in the early stage of breast cancinogenesis, but may play a role of in the progression of the cancer. RB gene methylation may also be a indicator in choice to identify the progression and prognosis of breast cancer.

Survivin: a promising biomarker in breast carcinoma

The antiapoptotic protein survivin can be detected in most types of malignant tumors, but it is rarely expressed in corresponding normal adult tissues. Therefore, survivin appears to represent a promising diagnostic biomarker. We examined survivin expression in 13 cases of normal breast tissue, 38 cases of fibroadenomas and 80 cases of breast carcinomas by immunohistochemical staining using anti-survivin antibody (DAKO, Clone 12C4). In each section, the intensity of staining, percentage of labeled cells, and the subcellular location of survivin antigen were assessed. Survivin was detected in 4/13 cases of normal breast tissue (30.7%), in 28/38 cases of fibroadenomas (73.7%), and in 67/80 cases of carcinomas (83.8%). Normal breast tissue showed cytoplasmic positivity only. In fibroadenomas, 19 cases (50.0%) revealed cytoplasmic reaction, and in 9 cases (23.7%), small foci of cells with combined nuclear and cytoplasmic location were identified. In carcinomas, cytoplasmic staining was found in 12/80 cases (15.0%), nuclear staining in 10/80 cases (12.5%), and combined cytoplasmic and nuclear staining in 45/80 cases (56.3%). Subcellular location of survivin between benign and malignant lesions revealed significant differences (p<0,001). Our findings point at practical use of survivin detection. We confirm the importance of nuclear staining of survivin antigen in breast carcinoma, which seems to be a notable diagnostic marker for estimation of the degree of neoplasia.

Expression of Twist, E-cadherin and N-cadherin in breast carcinoma and their clinical significance

To investigate the expression of Twist, E-cadherin and N-cadherin in breast carcinoma tissue and to analyse their effects on the breast carcinoma differentiation, size, infiltration and metastasis of the breast carcinoma. The expression of Twist, E-cadherin and N-cadherin in 56 cases of breast invasive ductal carcinoma, 38 cases of invasive lobular carcinoma, 41 cases of carcinoma in situ and 10 cases of normal breast tissue was detected using immunohistochemistry. (1) The expression rate of Twist in three types of breast carcinoma was 46.4% (26/56), 79.0% (30/38) and 26.8% (11/41) respectively, and the expression of Twist in invasive lobular carcinoma was significantly higher than that in invasive ductal carcinoma and carcinoma in situ (P = 0.002, P = 0.000). The expression rate of E-cadherin in three types of breast carcinoma was 78.6% (44/56), 29.0% (11/38) and 80.5% (33/41) respectively, and the expression of E-cadherin in invasive ductal carcinoma and carcinoma in situ was significantly higher than that in invasive lobular carcinoma (P = 0.000, P = 0.000). The expression rate of N-cadherin in three types of breast carcinoma was 53.6% (30/56), 68.4% (26/38) and 31.7% (13/41) respectively, and the expression of N-cadherin in invasive ductal carcinoma and invasive lobular carcinoma was significantly higher than that in carcinoma in situ (P = 0.033, P = 0.001). (2) In all the 135 cases, the expression of Twist was not correlated with that of E-cadherin (P = 0.005, Spearman correlation coefficient = -0.239), however, there was a positive correlation between the expression of Twist and N-cadherin and statistically significant(P = 0.000, Spearman correlation coefficient = 0.319). (3) In the invasive ductal carcinoma, the expression of N-cadherin in poorly-differentiated carcinoma was significantly higher than that of the moderately-or well-differentiated ones (P = 0.004). (4) In the invasive lobular carcinoma, the expression of Twist in cases with lymph node metastasis was significantly higher than that of cases without metastasis (P = 0.037). Twist, E-cadherin and N-cadherin have different expression patterns in the three kinds of breast carcinoma. The positive expression of Twist was correlated to lymph node metastasis in invasive lobular carcinoma and the positive expression of N-cadherin was correlated to cell the tissue differentiation in invasive ductal carcinoma. Detection of the expression of these biomarkers may provide a valuable reference for the study of breast carcinoma progression, metastasis and for the judgment of the biological behavior of the carcinoma.

Immunohistochemical expression of mTOR protein in breast carcinoma tissues

Immunohistochemical expression of mTOR protein in breast carcinoma tissues

The expression and clinical significance of p130Cas and c-erbB-2 in human breast cardnoma

Objective To explore the expression of p130Cas and c-erbB-2 in human breast carcinoma and the relationship between p130Cas and c-erbB-2 levels with clinical and pathological characteristics. Methods Immunohistochemistry SP staining was applied to detect the expression of p130Cas and c-erbB-2 in tumor tissues from 53 cases of human primary breast carcinoma,10 cases of breast fibroadenoma and in 10 cases of normal breast tissues. Results Breast carcinoma tissues showed higher levels of p130Cas and c-erbB-2 than normal and fibroadenoma tissues(both P＜0.01).The expression of p130Cas was related with age, menopausal status, ER, PR status and histological grades, but not related with tumor size, lymph node status and pathological stages. The expression of c-erbB-2 was related with ER status, histological grades and lymph node status, but not related with age, menopausal status, PR status, tumor size and pathological stages. There were no significant correlations between p130Cas and c-erbB-2. Conclusion p130Cas and c-erbB-2 have relations with the malignant transformation and differentiation of breast carcinoma.c-erbB-2 is associated with metastasis of breast carcinoma.p130Cas and c-erbB-2 can be served as useful indicators in the prognosis of breast carcinoma. Key words: Breast neoplasms; Genes,erbB-2; Adaptor proteins,signal trandueing

Expression of CD44s, CD44v6, and Hyaluronan Across the Spectrum of Normal-hyperplasia-carcinoma in Breast

The interaction between transmembrane receptors on epithelial tumor cells and the surrounding extracellular matrix molecules is important in tumor progression and metastasis. This interaction is best exemplified by the relationship of the receptor CD44 and the extracellular matrix component hyaluronan (HA). This study seeks to evaluate the expression and the correlation of CD44s, CD44v6, and HA in normal, hyperplastic, and malignant breast epithelium and stroma. Archival paraffin-embedded tissue from cases of normal breast tissue (n=10), intraductal hyperplasia without atypia (n=13), ductal carcinoma in situ (DCIS) (n=24), stage I infiltrating ductal carcinoma (n=28), stage II infiltrating ductal carcinoma (n=31), and their corresponding positive lymph nodes were retrieved from the surgical pathology files. Tissue sections were evaluated for the expression of CD44s, CD44v6, and HA in the epithelial and stromal cells by immunohistochemistry. Ductal epithelial cells and myoepithelial cells expressed CD44s in all cases of normal and benign breast tissue. The expression of CD44s in breast epithelium progressively decreased with increasing deviation from normal histology: 83% in DCIS, 46% in stage I ductal carcinoma and 26% in stage II ductal carcinoma. The reverse trend was observed for CD44v6 in ductal epithelium: 0% in normal breast, 15% in intraductal hyperplasia, 100% in DCIS, 82% in stage I infiltrating ductal carcinoma, 94% in stage II carcinoma, and 100% of metastatic carcinoma in the lymph nodes. HA was noted exclusively in the stroma but not in the epithelial cells. HA was faintly expressed in the intralobular stroma of normal breast tissue, confined to a narrow faint band adjacent to intraductal hyperplasia and localized to a broad well-defined band around DCIS. Stromal HA staining was more diffuse and intense in infiltrating carcinomas and was particularly pronounced surrounding the metastatic deposits in lymph nodes. This study demonstrates decreased expression of CD44s accompanied by increased expression of CD44v6 and increased stromal HA in breast cancer. These findings suggest that CD44s, CD44v6, and HA play complementary roles in the development and progression of breast cancer.

Significant increase of glucose transport activity in breast cancer

To study the expression level and significance of glucose transporter 1 (Glut-1) in normal breast tissue, adenosis, adenoma and breast carcinoma. A total of 147 cases of female breast tissue samples, including 92 cases of invasive ductal carcinoma, 26 cases of breast fibroadenoma, 24 cases of breast adenosis and 5 cases of normal breast tissues, were collected for quantitative detection of the expression of Glut-1 protein by immunohistochemistry (EnVision method) and Western blot, and its mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR). In normal breast tissue and benign lesions of the breast, Glut-1 was undetectable or only weakly detectable in cytoplasm of ductal and acinar epithelia. In contrast, the intensity of Glut-1 staining was significantly higher in invasive ductal carcinomas (P = 0.0002) with protein expression predominantly in cellular membrane and lesser in cytoplasm. Western blot and RT-PCR analyses showed that the expression of Glut-1 protein and mRNA were significantly increased in invasive ductal carcinoma than fibroadenoma (P =0.001 for protein; P <0.05 for mRNA) and adenosis (P =0.001 for protein; P < 0.05 for mRNA). There was a significant difference among groups (P = 0.0002 for protein; P = 0.0001 for mRNA). Glucose transport activity, as indicated by Glut-1 protein and its mRNA expression, significantly increases in breast carcinoma than non-cancerous lesions. The over-expression of Glut-1 in breast carcinoma is tightly coupled with tumor cell proliferation, invasion and metastasis, implying that Glut-1 may serve as a new marker in the early diagnosis and prognostication of breast malignancy as well as a new therapeutic target.

Raman spectroscopic analysis of breast cancer tissues: identifying differences between normal, invasive ductal carcinoma and ductal carcinoma in situ of the breast tissue

AbstractA relatively non‐destructive method employing Raman spectroscopy for the analysis of histopathological specimens is described. Raman spectroscopy has allowed qualitative analysis of the same specimen used for histopathological evaluation. Breast cancer tissues have been analysed to demonstrate the feasibility of the chemical changes taking place in the biological tissue, which can be identified precisely, and the results are reproducible.Raman analysis of tissue sections provides distinct spectra that can be used to distinguish between the nuclear grades of ductal carcinoma in situ (DCIS) and invasive ductal carcinoma (IDC) of the breast. Sixty cases of breast carcinoma including DCIS and IDC and seven cases of normal breast tissues were studied employing the Raman spectroscopic technique.This study reports for the first time spectral differences between DCIS grades. It is concluded that Raman spectroscopy can objectively distinguish between DCIS and IDC grades and is non‐destructive and reproducible. It should become possible in future to use Raman spectroscopy as an informative and quantitative method suitable for classification of grades and diagnosis of breast carcinoma. Copyright © 2007 John Wiley &amp; Sons, Ltd.

Expression of a cancer associated isoform of PCNA in breast cancer has implications as a potential biomarker

21069 Background: We have identified a novel cancer associated isoform of the protein, proliferating cell nuclear antigen (PCNA), termed (caPCNA), and demonstrated that this isoform arises through a direct protein post-translational modification, and not via genetic mutation or RNA splice variation. Our results suggest that caPCNA has the potential to serve as a highly effective and unique marker for identifying malignant breast cancer cells. Methods: This assertion is based on our proteomic-based analyses of more than 60 malignant and non- malignant breast cell lines and tissues. Commercially available antibodies against PCNA cannot distinguish between the different isoforms of PCNA present in malignant and non-malignant breast cells and cannot be used clinically to differentiate between normal and malignant breast tissue. However, we have recently developed a rabbit polyclonal antibody, (caPCNAab), which specifically recognizes only the caPCNA isoform expressed by malignant human breast cells. Results: Using this antibody we clearly show that caPCNA is expressed only in malignant breast cells and tissues, and can be found in early disease. caPCNA expression in tissues was quantified as average staining intensity X average percentage of cells stained. Using this criterion the following data were obtained: 10 cases of normal breast tissue (reduction mammoplasty) gave a total score of 1%; 35 cases of normal breast tissue adjacent to malignancy scored as 4%; 30 cases of DCIS scored as 90%; and 55 cases of invasive breast carcinoma scored as 120%. The five cases of ADH examined thus far were shown to score similar to that of normal breast tissue. Conclusions: The implication of these data is that the development of a caPCNAab-based IHC stain could potentially be used to reliably stain only in situ or invasive carcinoma, and distinguish genuinely benign lesions (e.g., ADH) from carcinoma, (e.g., DCIS) allowing definitive diagnosis in such cases where a limited amount of an atypical lesion prevents definitive diagnosis on routine H/E stained sections alone. For the patient population, this could result in a marked decrease in the need for either a repeat core biopsy or an excisional biopsy due to an inconclusive initial diagnosis. No significant financial relationships to disclose.

Expression of PH20 in primary and metastatic breast cancer and its pathological significance

To evaluate the expression pattern of PH20 in primary and metastatic breast cancer and its relationship to tumor metastatic potential. Anti-PH20 antibody was synthesized by injection of conjugated human PH20 peptides into rabbits. Immunohistochemical study was performed on 53 cases of human breast cancer. Western blot was used to detect PH20 expression in 5 cases of breast cancer with available fresh tissue. Two oligonucleotide probes were prepared for in-situ hybridization using breast tissue microarray. Normal breast tissue did not express PH20 (0/3), while 58.4% (31/53) of breast cancer cases did. The highest expression rate was found in metastatic foci in regional lymph nodes (83.3%), followed by primary breast cancer tissue in cases with lymph node secondaries (70.8%). The breast cancer cases with no any metastasis had an expression rate of 48.2%. The immunohistochemical staining results were further confirmed by Western blotting. In-situ hybridization showed PH20 RNA in 75% of the breast cancer tissue (21/28). Two of the 17 cases of normal breast tissue showed weak expression in some ductolobular units. The expression of PH20 has a positive correlation with metastatic potential in breast cancer. It is possible that PH20 may play an important role in the invasive growth and metastasis of breast cancer cells, via mechanisms such as digestion of surrounding stromal tissue and release of FGF-2.

Bcl-2 expression in the spectrum of preinvasive breast lesions.

The bcl-2 gene encodes for a protein that blocks apoptosis. Although the bcl-2 protein has been identified in some invasive breast cancers, its expression in preinvasive breast lesions has not been well characterized. We studied bcl-2 expression using immunohistochemistry in cases of normal breast tissue (n = 10), ductal hyperplasia (n = 18), atypical ductal hyperplasia (ADH) (n = 10), atypical lobular hyperplasia/lobular carcinoma in situ (lobular neoplasia, LN) (n = 10), and ductal carcinoma in situ (DCIS) (n = 42). We also related bcl-2 expression to p53 expression in these lesions because the p53 gene is altered in many invasive breast cancers and is also involved in the regulation of apoptosis. bcl-2 was consistently expressed in the epithelial cells in all normal breast tissue, ductal hyperplasia, ADH, and LN lesions. In contrast, bcl-2 expression was present in 76% of the DCIS cases and was related to the histologic grade of DCIS. Staining for bcl-2 was observed in 100% of the well differentiated DCIS cases, 90% of intermediately differentiated cases, and 33% of poorly differentiated cases (P < 0.001). No immunoreactivity for the p53 protein was seen in any of the cases of normal breast tissue, ductal hyperplasia, ADH, or LN lesions. However, 24% of the DCIS cases were p53 positive. The bcl-2+/p53- phenotype, as seen in all cases of normal breast tissue, ductal hyperplasia, ADH, and LN was also observed in 67% of the DCIS cases. In contrast, the remaining 33% of DCIS cases showed combinations of bcl-2 and p53 expression that differed from that of normal breast epithelium and the other pathologic lesions studied. Most lesions with altered bcl-2/p53 phenotypes, including all bcl-2-/p53+ cases, represented examples of poorly differentiated DCIS. The bcl-2 protein is consistently expressed in normal breast epithelium, ductal hyperplasia, ADH, and LN. bcl-2 expression is variable in DCIS. Among DCIS cases, bcl-2 is most common in well differentiated and intermediately differentiated lesions. Most DCIS lesions are bcl-2+ and p53-, similar to normal epithelium, benign proliferative lesions, and LN. However, a minority of DCIS lesions show combinations of bcl-2 and p53 expression that differ from normal breast epithelium and from the other pathologic lesions studied. It is possible that such lesions may represent a subset of DCIS in which the regulation of apoptosis is no longer under normal control mechanisms, resulting in enhanced tumor cell survival and tumor growth.

CDw75 Antigen Expression in Breast Lesions

Alterations on the cell surface of the oligosaccharide portion of glycoproteins and glycolipids are thought to play a role in tumorigenesis. Sialyltransferase catalyzes the incorporation of sialic acid to the carbohydrate group of glycoconjugates. Sialyltransferase has been found elevated in different tumour tissues and in the serum of cancer patients. In the present study we have examined the expression of the beta-galactosyl alpha 2,6-sialyltransferase requiring epitope CDw75, with the monoclonal antibody HH2. 142 breast lesions were included. 21% of the carcinomas in situ and 35% of the invasive carcinomas showed a diffuse cytoplasmic staining. Seven cases of invasive carcinomas also showed a distinct membrane immunoreactivity. We found no correlation between reactivity for CDw75 in malignant lesions and their metastatic potential. Only five out of 11 primary tumours with metastases expressed CDw75 in the primary tumour. In the benign lesions, there was a positive reaction in proliferating lesions, e.g. intraductal papillomas (2 out of 3 cases) and in epithelial proliferations in fibrocystic disease (10 out of 14 cases). None of the four fibroadenomas and phyllodes tumours and only one out of 22 cases of normal breast tissues showed immunoreactivity for HH2. In the malignant lesions, CDw75 was more frequently expressed in the carcinomas of high malignancy grade. The high frequency of immunoreactivity among the benign breast lesions can be indicative of activation of the epithelial cells.