Marie-Christine Mathieu

Evaluation of Tumor Angiogenesis of Breast Carcinoma Using Contrast-Enhanced Digital Mammography

OBJECTIVE The purpose of this article is to assess the accuracy of contrast-enhanced digital mammography in the detection of breast carcinoma and to correlate the findings on the images with those of histologic analysis using microvessel quantification. SUBJECTS AND METHODS Twenty patients with a suspicious breast abnormality underwent contrast-enhanced digital mammography using a full-field digital mammography unit that was modified to detect iodinated enhancement. For each patient, a total of six contrast-enhanced craniocaudal views were acquired from 30 seconds to 7 minutes after the injection of a bolus of 100 mL of an iodinated contrast agent. Image processing included a logarithmic subtraction and the analysis of enhancement kinetic curves. Contrast-enhanced digital mammography findings were compared with histologic analysis of surgical specimens, including intratumoral microvessel density quantification evaluated on CD34-immunostained histologic sections obtained from all patients. RESULTS An area of enhancement was depicted on contrast-enhanced digital mammograms in 16 of the 20 histologically proven breast carcinomas. Excellent correlation was seen between the size of enhancement and the histologic size of tumors, which ranged from 9 to 22 mm. Early enhancement with washout was observed in four cases, early enhancement followed by a plateau in four cases, gradual enhancement in seven cases, and unexpected decrease of enhancement in one case. Intratumoral microvessel density ranged from 11.7 to 216.6 microvessels per square millimeter. A poor correlation was found between data measured on contrast-enhanced digital mammography and intratumoral microvessel density measured on CD34-immunostained histologic sections. CONCLUSION Contrast-enhanced digital mammography is able to depict angiogenesis in breast carcinoma. Breast compression and projective images acquisition alter the quantitative assessment of enhancement parameters.

Lobular and ductal carcinomas of the breast have distinct genomic and expression profiles

Invasive ductal carcinomas (IDCs) and invasive lobular carcinomas (ILCs) are the two major pathological types of breast cancer. Epidemiological and histoclinical data suggest biological differences, but little is known about the molecular alterations involved in ILCs. We undertook a comparative large-scale study by both array-compared genomic hybridization and cDNA microarray of a set of 50 breast tumors (21 classic ILCs and 29 IDCs) selected on homogeneous histoclinical criteria. Results were validated on independent tumor sets, as well as by quantitative RT–PCR. ILCs and IDCs presented differences at both the genomic and expression levels with ILCs being less rearranged and heterogeneous than IDCs. Supervised analysis defined a 75-BACs signature discriminating accurately ILCs from IDCs. Expression profiles identified two subgroups of ILCs: typical ILCs (∼50%), which were homogeneous and displayed a normal-like molecular pattern, and atypical ILCs, more heterogeneous with features intermediate between ILCs and IDCs. Supervised analysis identified a 75-gene expression signature that discriminated ILCs from IDCs, with many genes involved in cell adhesion, motility, apoptosis, protein folding, extracellular matrix and protein phosphorylation. Although ILCs and IDCs share common alterations, our data show that ILCs and IDCs could be distinguished on the basis of their genomic and expression profiles suggesting that they evolve along distinct genetic pathways.

Radio-induced malignancies after breast cancer postoperative radiotherapy in patients with Li-Fraumeni syndrome

BackgroundThere are no specific recommendations for the management of breast cancer patients with germ-line p53 mutations, an exceptional genetic condition, particularly regarding postoperative radiotherapy. Preclinical data suggested that p53 mutations conferred enhanced radiosensitivity in vitro and in vivo and the few clinical observations showed that Li-Fraumeni families were at a higher risk of secondary radio-induced malignancies.MethodsWe reviewed a cohort of patients with germ-line p53 mutations who had been treated for breast cancer as the first tumor event. We assessed their outcome and the incidence of secondary radio-induced malignancies.ResultsAmong 47 documented Li-Fraumeni families treated from 1997 to 2007 at the Institut Gustave Roussy, 8 patients had been diagnosed with breast cancer as the first tumor event. Three patients had undergone conservative breast surgery followed by postoperative radiotherapy and five patients had undergone a mastectomy (3 with postoperative radiotherapy). Thus, 6/8 patients had received postoperative radiotherapy. Median follow-up was 6 years. Median age at the diagnosis of the primary breast cancer was 30 years. The histological characteristics were as follows: intraductal carcinoma in situ (n = 3), invasive ductal carcinoma (n = 4) and a phyllodes tumor (n = 1). Among the 6 patients who had received adjuvant radiotherapy, the following events had occurred: 3 ipsilateral breast recurrences, 3 contralateral breast cancers, 2 radio-induced cancers, and 3 new primaries (1 of which was an in-field thyroid cancer with atypical histology). In contrast, only one event had occurred (a contralateral breast cancer) among patients who had not received radiation therapy.ConclusionsThese observations could argue in favor of bilateral mastectomy and the avoidance of radiotherapy.

Epstein-Barr Virus (EBV) Genome and Expression in Breast Cancer Tissue: Effect of EBV Infection of Breast Cancer Cells on Resistance to Paclitaxel (Taxol)

ABSTRACT The Epstein-Barr virus (EBV) has been detected in subsets of breast cancers. In order to elaborate on these observations, we quantified by real-time PCR (Q-PCR) the EBV genome in biopsy specimens of breast cancer tissue as well as in tumor cells isolated by microdissection. Our findings show that EBV genomes can be detected by Q-PCR in about half of tumor specimens, usually in low copy numbers. However, we also found that the viral load is highly variable from tumor to tumor. Moreover, EBV genomes are heterogeneously distributed in morphologically identical tumor cells, with some clusters of isolated tumor cells containing relatively high genome numbers while other tumor cells isolated from the same specimen may be negative for EBV DNA. Using reverse transcription-PCR, we detected EBV gene transcripts: EBNA-1 in almost all of the EBV-positive tumors and RNA of the EBV oncoprotein LMP-1 in a smaller subset of the tissues analyzed. Moreover, BARF-1 RNA was detected in half of the cases studied. Furthermore, we observed that in vitro EBV infection of breast carcinoma cells confers resistance to paclitaxel (taxol) and provokes overexpression of a multidrug resistance gene (MDR1). Consequently, even if a small number of breast cancer cells are EBV infected, the impact of EBV infection on the efficiency of anticancer treatment might be of importance.

Breast cancer stroma frequently recruits fetal derived cells during pregnancy

IntroductionBreast carcinomas associated with pregnancy display a high frequency of inflammatory types, multifocal lesions and lymph node metastasis. Because pregnancy results in transfer to mothers of foetal stem cells that can migrate and differentiate into various tissues, we addressed the issue of whether such cells are present in breast carcinoma associated with pregnancy.MethodsWe analyzed women presenting with such tumours who were pregnant with male foetuses using fluorescence in situ hybridization (FISH), targeting X and Y chromosomes. The foetal cell phenotype was then determined by combining FISH and immunohistochemistry with various antibodies. Statistical analysis was performed using t-test or nonparametric Wilcoxons test.ResultsWe found that foetal cells were present in nine out of 10 carcinomas, in contrast with none of four benign mammary lesions (P < 0.05). Counting foetal and maternal cells showed that the mean number of foetal cells per million maternal cells was 36 in breast cancers and 0 in control samples (P < 0.01). By combining FISH and immunolabelling, we found that foetal cells expressed mainly mesenchymal or, to a lesser degree, epithelial or endothelial markers, but never leucocytes.ConclusionThese findings demonstrate the frequent presence of foetal derived cells essentially in tumour stroma. Given the role played by stroma in tumour proliferation, these findings raise the issue of whether foetal cell can be targeted to influence tumour behaviour.

Breast-conserving surgery after neoadjuvant anthracycline-based chemotherapy for large breast tumors

Randomized trials comparing neoadjuvant versus adjuvant chemotherapy show that primary chemotherapy allows more frequent breast‐preserving surgery even though no survival advantage has been demonstrated. The aim of the current study was to determine the predicting factors and the survival impact of breast conservation in patients with large breast tumors treated with neoadjuvant chemotherapy.

Immunohistochemical staining of bone marrow biopsies for detection of occult metastasis in breast cancer.

SummaryImmunohistochemical (IHC) techniques should allow for a greater detection of bone marrow micrometastasis in patients with breast carcinoma. We studied a series of bone marrow (BM) biopsies negative by conventional histologic techniques from 93 patients with breast carcinoma. Prior to this study, twelve BM biopsies, positive by conventional histology, were stained with a panel of monoclonal antibodies (MoAb), directed either against cytokeratin (KL1, AE1-AE3, CAM5-2) or epithelial membrane antigen (EMA, HMFG2). KL1 appeared to be the most sensitive of the markers used in the detection of metastases and is available commercially. It therefore was the only MoAb used with the series of 93 BM biopsies negative by conventional examination. Within this series, among 45 patients clinically suspected of having bone marrow metastasis but with BM biopsies negative by conventional staining, one case showing myelofibrosis stained positive with KL1 demonstrating isolated tumor cells. For the 48 patients without suspicion of bone marrow metastasis at initial diagnosis for breast carcinoma, KL1 revealed no marrow metastasis.Single bone marrow biopsy techniques whether stained by conventional or IHC methods do not appear to be useful tests to detect occult bone marrow metastasis, especially at initial diagnosis of clinically Mo breast carcinoma patients.

Surgical Clips Assist in the Visualization of the Lumpectomy Cavity in Three-Dimensional Conformal Accelerated Partial-Breast Irradiation

PURPOSE To determine to what extent the placement of surgical clips helps delineate the cavity in three-dimensional conformal accelerated partial-breast irradiation. PATIENTS AND METHODS Planning CT images of 100 lumpectomy cavities were reviewed in a cohort of 100 consecutive patients. The cavities were determined and categorized by two radiation oncologists according to cavity visualization score criteria and the breast density score. The two physicians first attempted to delineate the lumpectomy cavity without clips and then with clips. RESULTS In the case of high-density mammary tissue, the breast remodeling done during surgery does not enable the lumpectomy cavity to be sufficiently visualized. The use of surgical clips significantly improved the ability to visualize the lumpectomy cavity, with a 69% rate of concordance between physicians regardless of the breast tissue density. CONCLUSION The placement of surgical clips at lumpectomy enables visualization of the lumpectomy cavity and allows upgrading of the cavity visualization score on CT scans obtained for accelerated partial-breast irradiation treatment planning.

c-myc, p53 and bcl-2, apoptosis-related genes in infiltrating breast carcinomas: evidence of a link between bcl-2 protein over-expression and a lower risk of metastasis and death in operable patients.

Apoptosis is an important physiological process controlled by multiple genes, including c‐myc, p53 and bcl‐2, and its inhibition may lead to the development of human cancers. In this study, we analyzed expression of the c‐myc gene using Northern blot and of the p53 and bcl‐2 proteins by immuno‐histochemistry in 175 breast tumor specimens obtained from patients with operable breast cancer. We evaluated the relation between expression of these 3 genes and (i) the main usual prognostic factors (tumor size, histo‐prognostic grade, hormone receptors and number of positive nodes); (ii) the risk of death and relapse, taking into account these 4 factors, after a mean period of follow‐up of 9.5 years (SD 2 years). Over‐expression of c‐myc, p53 and bcl‐2 was observed in 35%, 23% and 63% of tumors, respectively. Over‐expression of c‐myc was strongly linked to the number of positive nodes (p = 0.0005). p53 protein expression was associated with both high‐grade (p = 0.0001) and hormone receptor‐negative (p = 0.0001) tumors. In contrast, bcl‐2 protein over‐expression was associated with the main favorable prognostic factors and, more particularly, with hormone receptor‐positive tumors (p = 0.0001). Multivariate analysis, using the Cox model, showed that only 2 factors were independently linked to the risk of death: number of positive nodes, which increased the risk (p = 0.0001), and bcl‐2 protein over‐expression, which decreased the risk (p = 0.008). When bcl‐2 over‐expression was studied in relation to nodal status, hormone receptor status and chemo‐ and hormone therapy, no significant difference was observed between different subgroups of patients. bcl‐2 expression was also associated with a significantly lower risk of distant metastasis (p = 0.04). In conclusion, bcl‐2 expression characterizes a particular phenotype of breast cancer with a favorable prognosis, and it may therefore be used as a marker of long‐term survival. Int. J. Cancer (Pred. Oncol.) 84:562–567, 1999.

c-erbB-2 (HER-2/neu) gene amplification is a better indicator of poor prognosis than protein over-expression in operable breast-cancer patients.

Our aim was to compare the prognostic value of c‐erbB‐2 gene amplification analyzed by Southern blot with that of protein (p185) over‐expression measured by immunohistochemistry in 172 patients with operable breast cancer (BC). Amplification and p185 over‐expression were found in 31 (18%) and 51 (30%) BCs, respectively. All but 1 of the tumors showed both amplification and over‐expression, while 21 (12%) tumors displayed over‐expression without amplification. The risk of death associated with c‐erbB‐2 gene amplification and p185 over‐expression was evaluated by multivariate analysis, taking into account tumor size, histoprognostic grade, hormone receptors and axillary node status. During a mean follow‐up of 9.5 (±2) years, node involvement (p < 0.001), c‐erbB‐2 gene amplification (p = 0.02) and negative hormone receptors (p = 0.02) were found to be independent prognostic indicators of the risk of death. Over‐expression of p185 with no amplification was not correlated with this risk. When the risk of death associated with c‐erbB‐2 amplification was studied according to chemo‐ and hormone therapy, no significant difference was observed between subgroups of subjects. Amplification was also associated (p = 0.02) with the risk of multifocal distant metastases (i.e., metastases detected concomitantly in at least 2 sites) and, thus, with BC aggressiveness. These data show the importance of c‐erbB‐2 gene amplification in predicting the long‐term outcome of patients and in selecting eligible patients for c‐erbB‐2–targeted therapies.