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Novel molecular markers of malignancy in histologically normal and benign breast.

Nasir A, Chen DT, Gruidl M, Henderson-Jackson EB, Venkataramu C, McCarthy SM, McBride HL, Harris E, Khakpour N, Yeatman TJ - Patholog Res Int (2011)

Bottom Line: We also demonstrated an increasing expression of TOP2A protein on an independent test set of HNB/benign/reductionmammoplasties, atypical-ductal-hyperplasia with and without synchronous breast cancer, DCIS and IDC tissues using a custom tissue microarray (TMA).In conclusion, TOP2A, MCM2, and BUB1B proteins are potential molecular biomarkers of malignancy in histologically normal and benign breast tissues.Larger-scale clinical validation studies are needed to further evaluate the clinical utility of these molecular biomarkers.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomic Pathology, Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.

ABSTRACT
To detect the molecular changes of malignancy in histologically normal breast (HNB) tissues, we recently developed a novel 117-gene-malignancy-signature. Here we report validation of our leading malignancy-risk-genes, topoisomerase-2-alpha (TOP2A), minichromosome-maintenance-protein-2 (MCM2) and "budding-uninhibited-by-benzimidazoles-1-homolog-beta" (BUB1B) at the protein level. Using our 117-gene malignancy-signature, we classified 18 fresh-frozen HNB tissues from 18 adult female breast cancer patients into HNB-tissues with low-grade (HNB-LGMA; N = 9) and high-grade molecular abnormality (HNB-HGMA; N = 9). Archival sections of additional HNB tissues from these patients, and invasive ductal carcinoma (IDC) tissues from six other patients were immunostained for these biomarkers. TOP2A/MCM2 expression was assessed as staining index (%) and BUB1B expression as H-scores (0-300). Increasing TOP2A, MCM2, and BUB1B protein expression from HNB-LGMA to HNB-HGMA tissues to IDCs validated our microarray-based molecular classification of HNB tissues by immunohistochemistry. We also demonstrated an increasing expression of TOP2A protein on an independent test set of HNB/benign/reductionmammoplasties, atypical-ductal-hyperplasia with and without synchronous breast cancer, DCIS and IDC tissues using a custom tissue microarray (TMA). In conclusion, TOP2A, MCM2, and BUB1B proteins are potential molecular biomarkers of malignancy in histologically normal and benign breast tissues. Larger-scale clinical validation studies are needed to further evaluate the clinical utility of these molecular biomarkers.

No MeSH data available.


Related in: MedlinePlus

Serial archival sections representative of   histologically normal breast tissues with high-grade  molecular abnormality stained for H&E, TOP2A, MCM2 and BUB1B proteins. (a) Portion of a TDLU from a histologically normal breast tissue with high-grade molecular abnormality (Case  22, specimen 1495). Serial sections showing the same TDLU as in (a) with distinct nuclear immunoreactivity for TOP2A (b) and MCM2 (c) in the epithelial cell nuclei, and diffuse cytoplasmic immunoreactivity (2+) for BUB1B protein (d) in the mammary epithelial cells. (IMPOX staining; original magnifications 400x).
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fig3: Serial archival sections representative of histologically normal breast tissues with high-grade molecular abnormality stained for H&E, TOP2A, MCM2 and BUB1B proteins. (a) Portion of a TDLU from a histologically normal breast tissue with high-grade molecular abnormality (Case  22, specimen 1495). Serial sections showing the same TDLU as in (a) with distinct nuclear immunoreactivity for TOP2A (b) and MCM2 (c) in the epithelial cell nuclei, and diffuse cytoplasmic immunoreactivity (2+) for BUB1B protein (d) in the mammary epithelial cells. (IMPOX staining; original magnifications 400x).

Mentions: The stained slides were evaluated by the breast pathologist on the study with extensive experience in immunohistochemistry (AN). Immunohistochemical staining for TOP2A and MCM2 was localized to the nuclei of the tumor cells and the normal breast epithelium, while the expression of BUB1B protein was localized to the cytoplasm of the tumor and normal breast epithelial cells. In order to calculate TOP2A and MCM2 nuclear staining indices in IDC tissue sections, up to 2000 tumor cells and in the case of histologically normal breast tissues (HNB-HGMA and HNB-LGMA) tissue sections up to 500 nonneoplastic breast epithelial cells were evaluated by absolute counting of positive (stained) and negative (unstained) cells in each section. TOP2A and MCM2 indices were recorded as per cent positive nuclei as previously described [13]. As outlined in the scheme published by Gonzalez et al. [14], these evaluations were made in the highest expression areas of the tumor and histologically normal breast tissues (Figures 2(b), 2(c), 3(b), 3(c), 4(b), and 4(c)).


Novel molecular markers of malignancy in histologically normal and benign breast.

Nasir A, Chen DT, Gruidl M, Henderson-Jackson EB, Venkataramu C, McCarthy SM, McBride HL, Harris E, Khakpour N, Yeatman TJ - Patholog Res Int (2011)

Serial archival sections representative of   histologically normal breast tissues with high-grade  molecular abnormality stained for H&E, TOP2A, MCM2 and BUB1B proteins. (a) Portion of a TDLU from a histologically normal breast tissue with high-grade molecular abnormality (Case  22, specimen 1495). Serial sections showing the same TDLU as in (a) with distinct nuclear immunoreactivity for TOP2A (b) and MCM2 (c) in the epithelial cell nuclei, and diffuse cytoplasmic immunoreactivity (2+) for BUB1B protein (d) in the mammary epithelial cells. (IMPOX staining; original magnifications 400x).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3140260&req=5

fig3: Serial archival sections representative of histologically normal breast tissues with high-grade molecular abnormality stained for H&E, TOP2A, MCM2 and BUB1B proteins. (a) Portion of a TDLU from a histologically normal breast tissue with high-grade molecular abnormality (Case  22, specimen 1495). Serial sections showing the same TDLU as in (a) with distinct nuclear immunoreactivity for TOP2A (b) and MCM2 (c) in the epithelial cell nuclei, and diffuse cytoplasmic immunoreactivity (2+) for BUB1B protein (d) in the mammary epithelial cells. (IMPOX staining; original magnifications 400x).
Mentions: The stained slides were evaluated by the breast pathologist on the study with extensive experience in immunohistochemistry (AN). Immunohistochemical staining for TOP2A and MCM2 was localized to the nuclei of the tumor cells and the normal breast epithelium, while the expression of BUB1B protein was localized to the cytoplasm of the tumor and normal breast epithelial cells. In order to calculate TOP2A and MCM2 nuclear staining indices in IDC tissue sections, up to 2000 tumor cells and in the case of histologically normal breast tissues (HNB-HGMA and HNB-LGMA) tissue sections up to 500 nonneoplastic breast epithelial cells were evaluated by absolute counting of positive (stained) and negative (unstained) cells in each section. TOP2A and MCM2 indices were recorded as per cent positive nuclei as previously described [13]. As outlined in the scheme published by Gonzalez et al. [14], these evaluations were made in the highest expression areas of the tumor and histologically normal breast tissues (Figures 2(b), 2(c), 3(b), 3(c), 4(b), and 4(c)).

Bottom Line: We also demonstrated an increasing expression of TOP2A protein on an independent test set of HNB/benign/reductionmammoplasties, atypical-ductal-hyperplasia with and without synchronous breast cancer, DCIS and IDC tissues using a custom tissue microarray (TMA).In conclusion, TOP2A, MCM2, and BUB1B proteins are potential molecular biomarkers of malignancy in histologically normal and benign breast tissues.Larger-scale clinical validation studies are needed to further evaluate the clinical utility of these molecular biomarkers.

View Article: PubMed Central - PubMed

Affiliation: Department of Anatomic Pathology, Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.

ABSTRACT
To detect the molecular changes of malignancy in histologically normal breast (HNB) tissues, we recently developed a novel 117-gene-malignancy-signature. Here we report validation of our leading malignancy-risk-genes, topoisomerase-2-alpha (TOP2A), minichromosome-maintenance-protein-2 (MCM2) and "budding-uninhibited-by-benzimidazoles-1-homolog-beta" (BUB1B) at the protein level. Using our 117-gene malignancy-signature, we classified 18 fresh-frozen HNB tissues from 18 adult female breast cancer patients into HNB-tissues with low-grade (HNB-LGMA; N = 9) and high-grade molecular abnormality (HNB-HGMA; N = 9). Archival sections of additional HNB tissues from these patients, and invasive ductal carcinoma (IDC) tissues from six other patients were immunostained for these biomarkers. TOP2A/MCM2 expression was assessed as staining index (%) and BUB1B expression as H-scores (0-300). Increasing TOP2A, MCM2, and BUB1B protein expression from HNB-LGMA to HNB-HGMA tissues to IDCs validated our microarray-based molecular classification of HNB tissues by immunohistochemistry. We also demonstrated an increasing expression of TOP2A protein on an independent test set of HNB/benign/reductionmammoplasties, atypical-ductal-hyperplasia with and without synchronous breast cancer, DCIS and IDC tissues using a custom tissue microarray (TMA). In conclusion, TOP2A, MCM2, and BUB1B proteins are potential molecular biomarkers of malignancy in histologically normal and benign breast tissues. Larger-scale clinical validation studies are needed to further evaluate the clinical utility of these molecular biomarkers.

No MeSH data available.


Related in: MedlinePlus