Different gene expression patterns in invasive lobular and ductal carcinomas of the breast.

Invasive ductal carcinoma (IDC) and invasive lobular carcinoma (ILC) are the two major histological types of breast cancer worldwide. Whereas IDC incidence has remained stable, ILC is the most rapidly increasing breast cancer phenotype in the United States and Western Europe. It is not clear whether IDC and ILC represent molecularly distinct entities and what genes might be involved in the development of these two phenotypes. We conducted comprehensive gene expression profiling studies to address these questions. Total RNA from 21 ILCs, 38 IDCs, two lymph node metastases, and three normal tissues were amplified and hybridized to approximately 42,000 clone cDNA microarrays. Data were analyzed using hierarchical clustering algorithms and statistical analyses that identify differentially expressed genes (significance analysis of microarrays) and minimal subsets of genes (prediction analysis for microarrays) that succinctly distinguish ILCs and IDCs. Eleven of 21 (52%) of the ILCs ("typical" ILCs) clustered together and displayed different gene expression profiles from IDCs, whereas the other ILCs ("ductal-like" ILCs) were distributed between different IDC subtypes. Many of the differentially expressed genes between ILCs and IDCs code for proteins involved in cell adhesion/motility, lipid/fatty acid transport and metabolism, immune/defense response, and electron transport. Many genes that distinguish typical and ductal-like ILCs are involved in regulation of cell growth and immune response. Our data strongly suggest that over half the ILCs differ from IDCs not only in histological and clinical features but also in global transcription programs. The remaining ILCs closely resemble IDCs in their transcription patterns. Further studies are needed to explore the differences between ILC molecular subtypes and to determine whether they require different therapeutic strategies.

Profile of immune cells in axillary lymph nodes predicts disease-free survival in breast cancer.

BACKGROUND: While lymph node metastasis is among the strongest predictors of disease-free and overall survival for patients with breast cancer, the immunological nature of tumor-draining lymph nodes is often ignored, and may provide additional prognostic information on clinical outcome. METHODS AND FINDINGS: We performed immunohistochemical analysis of 47 sentinel and 104 axillary (nonsentinel) nodes from 77 breast cancer patients with 5 y of follow-up to determine if alterations in CD4, CD8, and CD1a cell populations predict nodal metastasis or disease-free survival. Sentinel and axillary node CD4 and CD8 T cells were decreased in breast cancer patients compared to control nodes. CD1a dendritic cells were also diminished in sentinel and tumor-involved axillary nodes, but increased in tumor-free axillary nodes. Axillary node, but not sentinel node, CD4 T cell and dendritic cell populations were highly correlated with disease-free survival, independent of axillary metastasis. Immune profiling of ALN from a test set of 48 patients, applying CD4 T cell and CD1a dendritic cell population thresholds of CD4 > or = 7.0% and CD1a > or = 0.6%, determined from analysis of a learning set of 29 patients, provided significant risk stratification into favorable and unfavorable prognostic groups superior to clinicopathologic characteristics including tumor size, extent or size of nodal metastasis (CD4, p < 0.001 and CD1a, p < 0.001). Moreover, axillary node CD4 T cell and CD1a dendritic cell populations allowed more significant stratification of disease-free survival of patients with T1 (primary tumor size 2 cm or less) and T2 (5 cm or larger) tumors than all other patient characteristics. Finally, sentinel node immune profiles correlated primarily with the presence of infiltrating tumor cells, while axillary node immune profiles appeared largely independent of nodal metastases, raising the possibility that, within axillary lymph nodes, immune profile changes and nodal metastases represent independent processes. CONCLUSION: These findings demonstrate that the immune profile of tumor-draining lymph nodes is of novel biologic and clinical importance for patients with early stage breast cancer.

Ductal lavage of fluid-yielding and non-fluid-yielding ducts in BRCA1 and BRCA2 mutation carriers and other women at high inherited breast cancer risk.

OBJECTIVE: Nipple fluid production and atypical breast duct cells in women at high risk of breast cancer have been associated with further increased risk. Most publications on ductal lavage for cell collection report cannulating fluid-yielding ducts only. We report lavage of fluid-yielding and non-fluid-yielding ducts in women at high inherited breast cancer risk. METHODS: A pilot breast cancer screening study including ductal lavage was conducted in 75 women at high inherited risk, 56 (74.7%) of whom had BRCA1/2 mutations. Ductal lavage was attempted in any duct identifiable with a catheter. RESULTS: Ducts were successfully catheterized in 60 of 75 patients (80%). Successfully catheterized patients were younger (median age 41 versus 53 years, P = 0.0003) and more often premenopausal (51.7% versus 20%, P = 0.041). Thirty-one successfully catheterized patients [51.6%, 95% confidence interval (39.4-63.9%)] had non-fluid-yielding ducts only. Seventeen patients [28.3% (18.5-40.9%)] had atypical cells. Twelve of seventeen [70.6% (46.8-87.2%)] samples with atypia were from non-fluid-yielding ducts. Patients with non-fluid-yielding ducts (versus fluid-yielding ducts) were more likely to have had prior cancer (48.4% versus 17.2%, P = 0.014) or chemotherapy (45.2% versus 17.2%, P = 0.027); this was also true in patients with atypia from non-fluid-yielding ducts. CONCLUSION: Successfully lavaged women were younger and more often premenopausal. Atypical cells can be found in non-fluid-yielding ducts in patients at high inherited breast cancer risk. Non-fluid-yielding ducts, and atypia from non-fluid-yielding ducts, are more common in patients with prior cancer and chemotherapy. Larger studies are needed to identify risk factors and prognostic significance associated with atypia and non-fluid-yielding ducts in high-risk populations, and define their role as biomarkers.

Magnetic resonance imaging characteristics of fibrocystic change of the breast.

OBJECTIVE: The objective of this study was to identify magnetic resonance imaging (MRI) characteristics of fibrocystic change (FCC) of the breast. MATERIALS AND METHODS: Fourteen patients with a histopathologic diagnosis of solitary FCC of the breast underwent x-ray mammography and MRI of the breast. Three experienced breast imaging radiologists retrospectively reviewed the MRI findings and categorized the lesions on morphologic and kinetic criteria according to the ACR BI-RADS-MRI Lexicon. RESULTS: The most striking morphologic feature of fibrocystic change was nonmass-like regional enhancement found in 6 of 14 (43%) FCC lesions. Based on morphologic criteria alone, 12 of 14 (86%) lesions were correctly classified as benign. According to analysis of the time-intensity curves, 10 of 14 (71%) FCC lesions were correctly classified as benign. CONCLUSION: Although FCC has a wide spectrum of morphologic and kinetic features on MRI, it most often presents as a mass or a nonmass-like regional enhancing lesion with benign enhancement kinetics.

Rates of reexcision for breast cancer after magnetic resonance imaging-guided bracket wire localization.

BACKGROUND: We performed this study to determine rates of close or transected cancer margins after magnetic resonance imaging-guided bracket wire localization for nonpalpable breast lesions. STUDY DESIGN: Of 243 women undergoing MRI-guided wire localizations, 26 had MRI bracket wire localization to excise either a known cancer (n = 19) or a suspicious MRI-detected lesion (n = 7). We reviewed patient age, preoperative diagnosis, operative intent, mammographic breast density, MRI lesion size, MRI enhancement curve and morphology, MRI Breast Imaging Reporting and Data System (BI-RADS) assessment code, number of bracket wires, and pathology size. We analyzed these findings for their relationship to obtaining clear margins at first operative excision. RESULTS: Twenty-one of 26 (81%) patients had cancer. Of 21 patients with cancer, 12 (57%) had negative margins at first excision and 9 (43%) had close/transected margins. MRI size > or = 4 cm was associated with a higher reexcision rate (7 of 9, 78%) than those < 4 cm (2 of 12, 17%) (p = 0.009). MRI BI-RADS score, enhancement curve, morphology, and preoperative core biopsy demonstrating ductal carcinoma in situ (DCIS) were not predictive of reexcision. The average number of wires used for bracketing increased with lesion size, but was not associated with improved outcomes. On pathology, cancer size was smaller in patients with negative margins (12 patients, 1.2 cm) than in those with close/transected margins (9 patients, 4.6 cm) (p < 0.001). Reexcision was based on close/transected margins involving DCIS alone (6, 67%), infiltrating ductal carcinoma and DCIS (2, 22%), or infiltrating ductal carcinoma alone (1, 11%). Reexcision pathology demonstrated DCIS (3, 33%), no residual cancer (5, 55%), and 1 patient was lost to followup (1, 11%). Interestingly, cancer patients who required reexcision were younger (p = 0.022), but breast density was not associated with reexcision. CONCLUSIONS: To our knowledge, this is the first report of MRI-guided bracket wire localization. Patients with MRI-detected lesions less than 4 cm had clear margins at first excision; larger MRI-detected lesions were more likely to have close/transected margins. Reexcision was often because of DCIS and was the only pathology found at reexcision, perhaps because MRI is more sensitive for detecting invasive carcinoma than DCIS.

The Na+/I- symporter mediates iodide uptake in breast cancer metastases and can be selectively down-regulated in the thyroid.

PURPOSE: The Na(+)/I(-) symporter (NIS) is a key plasma membrane protein that mediates active iodide (I(-)) transport in the thyroid, lactating breast, and other tissues. Functional NIS expression in thyroid cancer accounts for the longstanding success of radioactive iodide ((131)I) ablation of metastases after thyroidectomy. Breast cancer is the only other cancer demonstrating endogenous functional NIS expression. Until now, NIS activity in breast cancer metastases (BCM) was unproven. EXPERIMENTAL DESIGN: Twenty-seven women were scanned with (99m)TcO(4)(-) or (123)I(-) to assess NIS activity in their metastases. An (131)I dosimetry study was offered to patients with I(-)-accumulating tumors. Selective down-regulation of thyroid NIS was tested in 13 patients with T(3) and in one case with T(3) + methimazole (MMI; blocks I(-) organification). NIS expression was evaluated in index and/or metastatic tumor samples by immunohistochemistry. RESULTS: I(-) uptake was noted in 25% of NIS-expressing tumors (two of eight). The remaining cases did not show NIS expression or activity. Thyroid I(-) uptakes were decreased to

Immunohistochemical profile of the sodium/iodide symporter in thyroid, breast, and other carcinomas using high density tissue microarrays and conventional sections.

Extrathyroidal cancers could potentially be targeted with (131)I, if the Na(+)/I(-) symporter (NIS) were functional. Using immunohistochemical methods we probed 1278 human samples with anti-NIS antibody, including 253 thyroid and 169 breast conventional whole tissue sections (CWTS). Four high density tissue microarrays containing a wide variety of breast lesions, normal tissues, and carcinoma cores were tested. The results of the normal microarray were corroborated in 50 CWTS. Nineteen of 34 normal tissues, including bladder, colon, endometrium, kidney, prostate, and pancreas, expressed NIS. Nineteen of 25 carcinomas demonstrated NIS immunopositivity; 55.7% of 479 carcinoma microarray cores expressed NIS, including prostate (74%), ovary (73%), lung (65%), colon (62.6%), and endometrium (56%). NIS protein was present in 75% benign thyroid lesions, 73% thyroid cancers, 30% normal-appearing, peritumoral breasts, 88% ductal carcinomas in situ, and 76% invasive breast carcinoma CWTS. Comparatively, breast microarray cores had lower immunoreactivity. Plasma membrane immunopositivity was confirmed in thyrocytes, salivary ductal, gastric mucosa, and lactating mammary cells. In other tissues, immunoreactivity was predominantly intracellular, particularly in malignant lesions. Thus, NIS is present in many normal epithelial tissues and is predominantly expressed intracellularly in many carcinomas. Elucidating the regulatory mechanisms that render NIS functional in extrathyroidal carcinomas may make (131)I therapy feasible.

Predictors of reexcision findings and recurrence after breast conservation.

PURPOSE: To identify predictors of reexcision findings and local recurrence in the setting of breast-conserving therapy with radiation. METHODS: The records of 535 patients who underwent breast-conserving surgery followed by radiation for Stage I or II cancer between 1972 and 1996 were reviewed. The mean follow-up period for surviving patients without evidence of recurrence is 6 years. Various clinical and pathologic prognostic factors were examined for significance with regard to reexcision findings and recurrence rates. Pathologic margin status was classified as negative, close (

Comparison of the comet assay and the oxygen microelectrode for measuring tumor oxygenation in head-and-neck cancer patients.

PURPOSE: To compare the Eppendorf PO2 histograph and the alkaline comet assay as methods of measuring tumor hypoxia in patients with head-and-neck squamous cell carcinomas. MATERIALS AND METHODS: As part of a larger clinical trial, 65 patients with head-and-neck squamous cell carcinoma nodal metastasis underwent tumor oxygenation measurements with Eppendorf PO2 histographs and comet assays, performed on fine-needle aspirates at 1 and 2 min after 5 Gy. Fifty-four patients had sufficient tumor cells for comet analysis at 1 min and 26 at both 1 and 2 min. Individual cells were examined for DNA single-strand breaks by alkaline gel electrophoresis, and the distribution of values was quantified using median tail moment (MTM). Nonirradiated tumor cells from pretreatment fine-needle aspirates received 5 Gy in vitro to establish the oxygenated response. RESULTS: There was a significant correlation between the 1- and 2-min MTM (slope = 0.77 +/- 0.03). There was no relationship between DNA damage in tumor cells irradiated in vitro and in vivo. No correlation was found between Eppendorf PO2 measurements and comet MTM. There was a statistically significant correlation between the treatment response in the node studied and comet MTMs, whereas no correlation was observed between treatment response and Eppendorf measurements. CONCLUSIONS: Comet assays are reproducible, as shown by biopsies at 1 and 2 min. Intertumor variation in the MTM is not a result of intrinsic radiosensitivity but of tumor hypoxia. There was no correlation between Eppendorf PO2 measurements and comet MTM. Comet assays were better than Eppendorf in predicting treatment response as an end point for short-term outcome. Longer follow-up is needed to determine the role of the comet assay as a predictor for locoregional tumor control and survivals.

Magnetic resonance imaging of intraductal papilloma of the breast.

To describe the appearance of isolated intraductal papilloma on contrast-enhanced water-specific, high spatial-resolution and rapid dynamic breast MRI, a retrospective review of unilateral breast images of 15 pathologically proven papilloma was performed. MRI revealed three patterns: Four papillomas were small, smooth, enhancing masses at the posterior end of an enlarged duct, corresponding to the "small lumenal mass" appearance of papilloma known from galactography. MRI detected two of these "small lumenal mass" papillomas in patients with abnormal nipple discharge even when galactography was unsuccessful. Seven papillomas were irregular enhancing masses detected in patients without nipple discharge. None of these papillomas had specifically benign findings. All seven demonstrated rapid enhancement and three showed rim enhancement or spiculation. These "tumor-like" papillomas mimicked invasive breast cancer on MRI. Four papillomas were occult on MRI, not revealed by either contrast-enhanced MRI or fat-suppressed T(2)-weighted MRI. Intraductal papillomas present with a variable appearance on MRI ranging from occult to "small lumenal mass" papillomas to irregular rapidly enhancing lesions that cannot be distinguished from invasive cancers.

Needle track seeding of papillary thyroid carcinoma from fine needle aspiration biopsy. A case report.

BACKGROUND: Dissemination of tumor cells from needle biopsy has been observed in a wide range of tumor types. Fine needle aspiration (FNA) biopsy has become accepted as the first-line test in the evaluation of thyroid nodules. Local recurrence of thyroid cancer from needle track seeding is an extremely rare complication of thyroid FNA. CASE: A 59-year-old woman developed local recurrence of papillary thyroid carcinoma three years after FNA of the primary cancer. Local metastases developed in the skin and sternocleidomastoid muscle. The location of the recurrent cancer and the linear relationship of the metastases indicated that local recurrence was due to needle track seeding at the time of FNA. CONCLUSION: Needle track seeding has been recognized as a possible, albeit rare, complication of FNA of thyroid cancer. Although proper FNA technique can reduce the potential for needle track seeding, its occurrence is an unavoidable complication of FNA evaluation of thyroid malignancies.

Bioluminescent monitoring of NIS-mediated (131)I ablative effects in MCF-7 xenografts.

Optical imaging has made it possible to monitor response to anticancer therapies in tumor xenografts. The concept of treating breast cancers with (131)I is predicated on the expression of the Na(+)/I- symporter (NIS) in many tumors and uptake of I- in some. The pattern of (131)I radioablative effects were investigated in an MCF-7 xenograft model dually transfected with firefly luciferase and NIS genes. On Day 16 after tumor cell implantation, 3 mCi of (131)I was injected. Bioluminescent imaging using d-luciferin and a cooled charge-coupled device camera was carried out on Days 1, 2, 3, 7, 10, 16, 22, 29, and 35. Tumor bioluminescence decreased in (131)I-treated tumors after Day 3 and reached a nadir on Day 22. Conversely, bioluminescence steadily increased in controls and was 3.85-fold higher than in treated tumors on Day 22. Bioluminescence in (131)I-treated tumors increased after Day 22, corresponding to tumor regrowth. By Day 35, treated tumors were smaller and accumulated 33% less (99m)TcO(4)(-) than untreated tumors. NIS immunoreactivity was present in <50% of (131)I-treated cells compared to 85-90% of controls. In summary, a pattern of tumor regression occurring over the first three weeks after (131)I administration was observed in NIS-expressing breast cancer xenografts.

Contrast-enhanced MRI of ductal carcinoma in situ: characteristics of a new intensity-modulated parametric mapping technique correlated with histopathologic findings.

PURPOSE: To identify morphologic and dynamic enhancement magnetic resonance imaging (MRI) features of pure ductal carcinoma in situ (DCIS) by using a new intensity-modulated parametric mapping technique, and to correlate the MRI features with histopathologic findings. MATERIALS AND METHODS: Fourteen patients with pure DCIS on pathology underwent conventional mammography and contrast-enhanced (CE) MRI using the intensity-modulated parametric mapping technique. The MR images were reviewed and the lesions were categorized according to morphologic and kinetic criteria from the ACR BI-RADS-MRI Lexicon, with BI-RADS 4 and 5 lesions classified as suspicious. RESULTS: With the use of a kinetic curve shape analysis, MRI classified seven of 14 lesions (50%) as suspicious, including four with initial-rapid/late-washout and three with initial-rapid/late-plateau. Using morphologic criteria, MRI classified 10/14 (71%) as suspicious, with the most prominent morphologic feature being a regional enhancement pattern. Using the intensity modulated parametric mapping technique, MRI classified 12/14 cases (86%) as suspicious. Parametric mapping identified all intermediate- and high-grade DCIS lesions. CONCLUSION: The intensity-modulated parametric mapping technique for breast MRI resulted in the highest detection rate for the DCIS cases. Furthermore, the parametric mapping technique identified all intermediate- and high-grade DCIS lesions, suggesting that a negative MRI using the parametric mapping technique may exclude intermediate- and high-grade DCIS. This finding has potential clinical implications.

MR imaging features of infiltrating lobular carcinoma of the breast: histopathologic correlation.

OBJECTIVE: Our study aimed to correlate the dynamic contrast-enhanced MR appearance of infiltrating lobular carcinoma of the breast with histopathologic findings. MATERIALS AND METHODS: We retrospectively reviewed the high-resolution, fat-suppressed and dynamic contrast-enhanced MR images of 13 of 20 women diagnosed with pathologically proven infiltrating lobular carcinoma of the breast. Twelve of the 13 women presented with breast symptoms and underwent mammography. Five of the women also had breast sonography. MR imaging was performed for evaluation of disease extent before the patients underwent modified radical mastectomy (n = 11) or lumpectomy (n = 2). Three experienced radiologists reviewed the MR scans. The tumor pattern types described on imaging were correlated with a detailed analysis of the pathology. RESULTS: We found three patterns of infiltrating lobular carcinoma on MR imaging. The tumor pattern on imaging correlated with pathologic tumor morphology. We found the following patterns of infiltrating lobular carcinoma: a solitary mass with irregular margins (n = 4) that corresponded to the same appearance at pathology; multiple lesions, either connected by enhancing strands (n = 6) or separated by nonenhancing intervening tissue (n = 2), that correlated with the pathologic appearance of noncontiguous tumor foci, with malignant cells streaming in single-file fashion in the breast stroma or small tumor aggregates separated by normal tissue; and enhancing septa only, which were correlated with the histopathologic appearance of tumor cells streaming in the breast stroma (n = 1). CONCLUSION: Infiltrating lobular carcinoma may be detected on MR imaging as solitary or multiple lesions that correspond to tumor morphology on pathologic examination. The appearance of multiple lesions or of enhancing fibroglandular breast elements on MR imaging is suggestive of infiltrating lobular carcinoma.

Breast magnetic resonance image screening and ductal lavage in women at high genetic risk for breast carcinoma.

BACKGROUND: Intensive screening is an alternative to prophylactic mastectomy in women at high risk for developing breast carcinoma. The current article reports preliminary results from a screening protocol using high-quality magnetic resonance imaging (MRI), ductal lavage (DL), clinical breast examination, and mammography to identify early malignancy and high-risk lesions in women at increased genetic risk of breast carcinoma. METHODS: Women with inherited BRCA1 or BRCA2 mutations or women with a >10% risk of developing breast carcinoma at 10 years, as estimated by the Claus model, were eligible. Patients were accrued from September 2001 to May 2003. Enrolled patients underwent biannual clinical breast examinations and annual mammography, breast MRI, and DL. RESULTS: Forty-one women underwent an initial screen. Fifteen of 41 enrolled women (36.6%) either had undergone previous bilateral oophorectomy and/or were on tamoxifen at the time of the initial screen. One patient who was a BRCA1 carrier had high-grade ductal carcinoma in situ (DCIS) that was screen detected by MRI but that was missed on mammography. High-risk lesions that were screen detected by MRI in three women included radial scars and atypical lobular hyperplasia. DL detected seven women with cellular atypia, including one woman who had a normal MRI and mammogram. CONCLUSIONS: Breast MRI identified high-grade DCIS and high-risk lesions that were missed by mammography. DL detected cytologic atypia in a high-risk cohort. A larger screening trial is needed to determine which subgroups of high-risk women will benefit and whether the identification of malignant and high-risk lesions at an early stage will impact breast carcinoma incidence and mortality.