Molecular characterization of CD44+/CD24-/Ck+/CD45- cells in benign and malignant breast lesions.

Breast cancer epithelial cells with the CD44+/CD24-/low phenotype possess tumor-initiating cells and epithelial-mesenchymal transition (EMT) capacity. Massive parallel sequencing can be an interesting approach to deepen the molecular characterization of these cells. We characterized CD44+/CD24-/cytokeratin(Ck)+/CD45- cells isolated through flow cytometry from 43 biopsy and 6 mastectomy samples harboring different benign and malignant breast lesions. The Ion Torrent Ampliseq Cancer Hotspot panel v2 (CHPv2) was used for the identification of somatic mutations in the DNA extracted from isolated CD44+/CD24-/Ck+/CD45- cells. E-Cadherin and vimentin immunohistochemistry was performed on sections from the corresponding formalin-fixed, paraffin-embedded (FFPE) blocks. The percentage of CD44+/CD24-/Ck+/CD45- cells increased significantly from non-malignant to malignant lesions and in association with a significant increase in the expression of vimentin. Non-malignant lesions harbored only a single-nucleotide polymorphism (SNP). Mutations in the tumor suppressor p53 (TP53), NOTCH homolog 1 (NOTCH1), phosphatase and tensin homolog (PTEN), and v-akt murine thymoma viral oncogene homolog 1 (AKT1) genes were found in isolated CD44+/CD24-/Ck+/CD45- cells from ductal carcinomas in situ (DCIS). Additional mutations in the colony-stimulating factor 1 receptor (CSF1R), ret proto-oncogene (RET), and TP53 genes were also identified in invasive ductal carcinomas (IDCs). The use of massive parallel sequencing technology for this type of application revealed to be extremely effective even when using small amounts of DNA extracted from a low number of cells. Additional studies are now required using larger cohorts to design an appropriate mutational profile for this phenotype.

Breast Density and Benign Breast Disease: Risk Assessment to Identify Women at High Risk of Breast Cancer.

PURPOSE: Women with proliferative breast lesions are candidates for primary prevention, but few risk models incorporate benign findings to assess breast cancer risk. We incorporated benign breast disease (BBD) diagnoses into the Breast Cancer Surveillance Consortium (BCSC) risk model, the only breast cancer risk assessment tool that uses breast density. METHODS: We developed and validated a competing-risk model using 2000 to 2010 SEER data for breast cancer incidence and 2010 vital statistics to adjust for the competing risk of death. We used Cox proportional hazards regression to estimate the relative hazards for age, race/ethnicity, family history of breast cancer, history of breast biopsy, BBD diagnoses, and breast density in the BCSC. RESULTS: We included 1,135,977 women age 35 to 74 years undergoing mammography with no history of breast cancer; 17% of the women had a prior breast biopsy. During a mean follow-up of 6.9 years, 17,908 women were diagnosed with invasive breast cancer. The BCSC BBD model slightly overpredicted risk (expected-to-observed ratio, 1.04; 95% CI, 1.03 to 1.06) and had modest discriminatory accuracy (area under the receiver operator characteristic curve, 0.665). Among women with proliferative findings, adding BBD to the model increased the proportion of women with an estimated 5-year risk of 3% or higher from 9.3% to 27.8% (P<.001). CONCLUSION: The BCSC BBD model accurately estimates women's risk for breast cancer using breast density and BBD diagnoses. Greater numbers of high-risk women eligible for primary prevention after BBD diagnosis are identified using the BCSC BBD model.

Categorization of non-mass-like breast lesions detected by MRI.

BACKGROUND: Breast MR imaging has emerged as a highly sensitive modality for the imaging of breast tumors. However, a standardized method of interpretation of lesions showing non-mass-like enhancement does not exist. The purpose of this study was to analyze the features of non-mass-like breast lesions detected by MRI, and to establish a standardized method of interpretation to allow categorization of these lesions. METHODS: A retrospective review was performed for 102 consecutive nonpalpable mammographically occult, non-mass-like lesions detected by MRI that had undergone ultrasound-guided vacuum-assisted biopsy. MR imaging was performed on a 1.5-Tesla system. The distribution patterns were classified into three categories as follows: single quadrant/solitary lesion (linear), single quadrant/grouped lesion (focal, regional, segmental), and multiquadrant lesion (multiple regions, diffuse). The presence of a ductal pattern was assessed in the enhancing lesions after the tumor distribution had been decided. In addition to the BI-RADS-MRI descriptors, the presence of clustered ring enhancement was also assessed in heterogeneous enhancing lesions. We divided non-mass-like lesions into those with a small (category 3a), moderate (category 3b), or substantial (category 4) likelihood of malignancy. RESULTS: The features with the highest positive predictive value (PPV) for cancer were clustered ring enhancement (67%) (P = 0.004), a branching-ductal pattern (38%) (P = 0.003), and clumped architecture (20%). The PPV for cancer of a linear-ductal pattern was 11% (1/9). All lesions showing multiquadrant distribution, linear-nonspecific lesion, non-branching pattern with homogeneous and stippled internal architectures, and heterogeneous lesion without clustered ring enhancement were diagnosed as benign. CONCLUSION: Non-mass-like breast lesions detected on MRI showing a clustered ring enhancement, a branching-ductal pattern, and clumped architecture should be evaluated further by biopsy (category 4), while lesions not showing these characteristics may be observed without unnecessary intervention (category 3a). Lesions showing a linear-ductal pattern may be followed carefully or evaluated by biopsy as needed (category 3b).

Immunophenotypic and prognostic analysis of E-cadherin and beta-catenin expression during breast carcinogenesis and tumour progression: a comparative study with CD44.

AIMS: This study was performed to investigate whether immunohistochemical expression of E-cadherin (E-cad) and beta-catenin (beta-cat) in conjunction with CD44 may correlate with the clinical evolution and prognosis of breast cancer. METHODS AND RESULTS: One-hundred and forty-two routinely processed breast tissue samples including normal breast, benign lesions, in situ and invasive carcinomas were investigated. E-cad and beta-cat were strongly expressed by luminal and basal cells in normal glands, benign proliferative and early neoplastic intraductal lesions. Contrarily, CD44 was expressed exclusively by myoepithelial cells in normal breast, whereas different isoform expression patterns were observed in premalignant and malignant lesions. Simultaneous lack of E-cad/beta-cat expression was detected in in situ and invasive lobular carcinomas in contrast to ductal lesions, in which the differential loss of the molecules was associated with poorer differentiation, irrespective of CD44 immunophenotype. Reduced E-cad (P = 0.003), beta-cat (P = 0.03) and increased CD44v4 (P = 0.005) and v7 (P = 0.007) expression were significantly associated with positive lymph node status. Decreased E-cad and lack of CD44v6 expression correlated with poor survival. There was no difference between the expression of either molecule in in situ and invasive components within the same tumour. CONCLUSIONS: Our results indicate that changes in E-cad, beta-cat and CD44 expression occur early in breast carcinogenesis; they are involved in tumour differentiation, but events additional to their deranged expression are needed to acquire an invasive phenotype.

Fine needle aspiration cytology of palpable breast lesions.

Fine needle aspiration cytology is a safe diagnostic technique for breast cancer and can identify certain benign lesions, which can be excised without affecting surrounding tissue. The combination of experienced operator and cytologist can yield high sensitivity and specificity, together with important prognostic information.

Breast disease. A clinical study with special reference to diagnostic procedures.

A consecutive series of 1.310 women with breast complaints and a control group of 241 healthy women were examined by clinical examination and mammography. The majority of the palpable and non-palpable lesions were preoperatively classified on a four-point scale according to the degree of suspicion of malignancy. A similar classification was performed after mammography as well as after aspiration biopsy. Some aspects of the case-histories including a positive family history of breast carcinoma, marital status and breast feeding were studied in women with breast complaints and compared to the findings in a control group of women. Previous breast disease, mode of detection, delay and breast symptoms were also studied...