Contrast-Enhanced Mammography-Guided Biopsy: Preliminary Results of a Single-Center Retrospective Experience.

Background: CEM-guided breast biopsy is an advanced diagnostic procedure that takes advantage of the ability of CEM to enhance suspicious breast lesions. The aim pf this paper is to describe a single-center retrospective experience on CEM-guided breast biopsy in terms of procedural features and histological outcomes. Methods: 69 patients underwent the procedure. Patient age, breast density, presentation, dimensions, and lesion target enhancement were recorded. All the biopsy procedures were performed using a 7- or 10-gauge (G) vacuum-assisted biopsy needle. The procedural approach (horizontal or vertical) and the decubitus of the patient (lateral or in a sitting position) were noted. Results: A total of 69 patients underwent a CEM-guided biopsy. Suspicious lesions presented as mass enhancement in 35% of cases and non-mass enhancement in 65% of cases. The median size of the target lesions was 20 mm. The median procedural time for each biopsy was 10 ± 4 min. The patients were placed in a lateral decubitus position in 52% of cases and seated in 48% of cases. The most common approach was horizontal (57%). The mean AGD was 14.8 mGy. At histology, cancer detection rate was 28% (20/71). Conclusions: CEM-guided biopsy was feasible, with high procedure success rates and high tolerance by the patients.

BOMB trial: First results of stereotactic radiotherapy to primary breast tumor in metastatic breast cancer patients.

Aim: A prospective dose escalation trial was developed to evaluate the maximum tolerated dose of stereotactic body radiotherapy (SABRT) to primary breast cancer in stage IV disease. The aim of the present report was to describe safety and outcome of the first dose level cohort of patients. Material and methods: Patients with histologically confirmed diagnosis of invasive breast carcinoma (biological immuno-histochemical profile: luminal and/or HER2 positive) and distant metastatic disease not progressing after 6 months of systemic therapy with a tumor CT or 5FDG-PET detectable were deemed eligible. The starting dose was 40 Gy in 5 fractions (level 1) because this dose proved to be safe in previous dose-escalation trial on adjuvant stereotactic body radiotherapy. The maximum dose level was chosen as 45 Gy in 5 fractions. Dose limiting toxicity was any grade 3 or worse toxicity according to CTCAE v.4. Time-to-event Keyboard (TITE-Keyboard) design (Lin and Yuan, Biostatistics 2019) was used to find the maximum tolerated dose (MTD). MTD was the dose of radiotherapy associated with a ≤ 20% rate pre-specified treatment-related dose-limiting toxicity (DLT). Results: To date 10 patients have been treated at the starting dose level. Median age was 80 years (range 50-89). 7 patients had a luminal disease, while 3 patients had an HER2 positive disease. No patient suspended ongoing systemic treatment. No protocol defined DLTs were observed. Grade 2 skin toxicity occurred in 4 patients with diseases located close to or involving the skin. Median follow-up was 13 months and all 10 patients were evaluable for response: 5 achieved a complete response, 3 achieved a partial response and 2 showed a stable disease, all with a clinical benefit (resolution of skin retraction, bleeding and pain). The mean reduction in the sum of the largest diameters of target lesions was of 61.4% (DS=17.0%). Conclusions: SABR to primary breast cancer seems feasible and is associated with symptoms reduction. Continued accrual to this study is needed to confirm the safety and assess the MTD. Clinical trial registration: ClinicalTrials.gov, identifier NCT05229575.

Subaxillary Replacement Flap Compared with the Round Block Displacement Technique in Oncoplastic Breast Conserving Surgery: Functional Outcomes of a Feasible One Stage Reconstruction.

BACKGROUND: For selected women diagnosed with breast cancer (BC), partial reconstructive techniques involve displacement or replacement procedures to improve cosmesis without compromising oncological safety. This study aims to evaluate the surgical outcomes of the round block (RB) compared with the subaxillary flap (SF) technique for patients with upper outer tumor. PATIENTS AND METHODS: Thirty-three patients treated with oncoplastic conserving surgery (15 RB and 18 SF) were enrolled in this retrospective study. After carrying out a comparison of baseline characteristics, all cases were recruited for postoperative evaluation of oncological and cosmetic parameters. Moreover, we investigated several scoring combinations to check whether they could discriminate surgeon and patient satisfaction according to different functional results. RESULTS: Median age (p < 0.05), average tumor size (p > 0.05), estimated resection volume (p > 0.05), and nodal involvement (p > 0.05) were slightly higher in the SF group. A greater frequency of DCIS (p < 0.05) in the RB series correlated with reintervention for positive margins (p > 0.001). At a mean follow-up of 19 months, no locoregional recurrences were recorded and early and late complications were comparable (p > 0.05). The overall satisfaction with cosmesis was characterized by similar proportions of good results (p > 0.05), with some details more related to each procedure. CONCLUSION: The proposed techniques represent effective solutions for reshaping that follows upper outer wide excision, achieving comparable complication rates, low reinterventions, and good aesthetic results in relation to technical and social functioning evaluations. However, it is crucial to establish a careful patient selection in order to manage correct surgical planning while predicting any potential sequelae or complication.

A Case of Secretory Carcinoma in a Patient With a History of Contralateral Medullary Carcinoma.

Background: Secretory and medullary carcinomas of the breast are rare subtypes of infiltrating ductal carcinoma. The different histological behavior of medullary and secretory carcinomas is correlated with different imaging features on mammography, ultrasound, and magnetic resonance imaging. Case Report: We report the case of a Caucasian woman in which both subtypes of tumors were diagnosed in an 8-year time interval and evaluate, in antithesis, histopathological and imaging aspects of medullary and secretory carcinoma. Conclusion: To our knowledge, this is the first case reported in literature of secretory carcinoma with a complete imaging tumor evaluation in a patient with a previous contralateral medullary cancer.

Preoperative Ultrasound-guided Sub-areolar Biopsy in Predicting Occult Nipple Involvement in Breast Cancer Patients: Proposal for a Methodological Approach.

BACKGROUND/AIM: The prediction of a sub-areolar tissue infiltration in breast cancer (BC) patients could be helpful in selecting the best functional outcome according to several reconstructive oncoplastic or radical techniques. This study aims to evaluate the diagnostic performance of preoperative ultrasound (US) guided sub-areolar biopsy (SAB) in detecting occult nipple involvement, in comparison with the definitive pathological examination of tissue after surgery. PATIENTS AND METHODS: We prospectively recorded clinical and pathological data of 46 consecutive patients scheduled for breast conserving surgery or nipple-areola sparing mastectomy. All cases underwent preoperative SAB and the results were compared with the histopathology of the dissected tumors and their biological characteristics. All parameters were correlated with nipple involvement by univariate and multivariate analysis. RESULTS: The sensitivity, specificity, overall accuracy, positive predictive value, and negative predictive value of the SAB examination for nipple staging were 60%, 97.5%, 75% and 95.2%, respectively. According to the clinicopathological features, the bivariate analysis did not show a significant interaction between patient age, tumor size or type, lymph node status, lymphatic vascular invasion, histologic grade, ER, PR, Ki-67 status, HER2 amplification, multifocal or multicentric disease and positive NAC assessment (p>0.05). CONCLUSION: A preoperative sample of retroareolar tissue obtained by US-guided biopsy could be considered a reproducible, mini-invasive diagnostic procedure useful to facilitate immediate breast reconstruction with implants or through conservative oncoplastic approaches, thereby guiding clinical practice.

Update on oncoplastic techniques in breast conserving surgery: algorithms for predictable results and custom-made reconstructions.

Breast cancer surgery is going to be a relevant specialty involved in the multidisciplinary disease management, finalized to guarantee high performances on mortality reduction together with a quality-of-life improvement. The intellectual architecture of this field is characterized by its inclination to respect, sharing, empathy and scientific knowledge that can make it a high model of medical culture. In this context, oncoplastic breast conserving surgery is identified by special aesthetic sensitivities, as well as creativity and innovation. Knowing how to reshape the gland and fill the gaps, planning the targeted exeresis design, while anticipating the final outcome, respecting symmetries and ensuring first of all oncological safety, are the new skills that are required to the breast surgeon. Several tecniques could provide a refined surgical treatment but always challenging according to anatomical findings and patient expectations, as a systematic process of self-critical reflection, with the purpose to optimize criteria for improving therapeutic results.

Sex Hormones Related Ocular Dryness in Breast Cancer Women.

BACKGROUND: Dry eye syndrome (DES) is strictly connected to systemic and topical sex hormones. Breast cancer treatment, the subsequent hormonal therapy, the subsequent hyperandrogenism and the early sudden menopause, may be responsible for ocular surface system failure and its clinical manifestation as dry eye disease. This local dryness is part of the breast cancer iatrogenic dryness, which affects overall mucosal tissue in the fragile population of those with breast cancer. METHODS: A literature review regarding the role of sex hormone changes and systemic hormonal replacement treatments (SHRT) in DES available on PubMed and Web of Science was made without any restriction of language. RESULTS: Androgens exert their role on the ocular surface supporting meibomian gland function and exerting a pro-sebaceous effect. Estrogen seems to show a pro/inflammatory role on the ocular surface, while SHRT effects on dry eye are still not well defined, determining apparently contradictory consequences on the ocular surface homeostasis. The role of sex hormones on dry eye pathogenesis is most likely the result of a strict crosstalk between the protective androgens effects and the androgen-modulating effects of estrogens on the meibomian glands. CONCLUSIONS: Patients with a pathological or iatrogenic hormonal imbalance, such as in the case of breast cancer, should be assessed for dry eye disease, as well as systemic dryness, in order to restore their social and personal quality of life.

Clinical Outcomes Among Major Breast Cancer Subtypes After Neoadjuvant Chemotherapy: Impact on Breast Cancer Recurrence and Survival.

BACKGROUND/AIM: Prior studies have underlined the prognostic relevance of pathological complete response (pCR) after neoadjuvant chemotherapy (NAC) in breast cancer. However, an accurate demonstration of treatment efficacy is dependent on its potential to predict long-term outcomes of recurrence and death, and this issue remains somewhat controversial. PATIENTS AND METHODS: One hundred and sixty-nine patients with breast cancer (BC) treated with NAC followed by surgery were enrolled in this retrospective study. After carrying out multivariable analyses, involving baseline characteristics (tumor stage, nodal status, histological grade, biological profile) and response status, we analysed the association between pCR and disease-free (DFS) and overall survival (OS) in various subtypes. Moreover, we investigated several residual disease-scoring combinations to check whether they could discriminate prognostic subsets according to their variable tumor range after NAC. RESULTS: Overall, factors associated with pCR were non-luminal subtype (p<0.001), high grade (p=0.001) and HER2-overexpression (p=0.001). Residual tumor and nodal stage after NAC significantly correlated with DFS (p=0.007) and OS (p<0.001). Similarly, pCR after NAC showed significantly better DFS (p=0.01), particularly for HER2-positive (p=0.003), triple-negative (p=0.019) and HER2-positive Luminal B profiles (p=0.019). However, there was no statistical difference in the OS among patients who had PCR, compared to absence of pCR (p=0.40). CONCLUSION: Extent of residual disease and evidence of regression provide helpful prognostic details in BC patients treated with NAC. Achieving pCR after NAC is related with significantly better DFS, with the potential of maximized breast and axillary conservation based on clinical response. The distribution of expertise in a cross-disciplinary setting could provide safe and favourable prognosis, while improving cosmetic outcomes and quality of life.

Delay in Breast Cancer Treatments During the First COVID-19 Lockdown. A Multicentric Analysis of 432 Patients.

BACKGROUND/AIM: Extraordinary restrictions aimed to limit Sars-CoV-2 spreading; they imposed a total reorganization of the health-system. Oncological treatments experienced a significant slowdown. The aim of our multicentric retrospective study was to evaluate screening suspension and surgical treatment delay during COVID-19 and the impact on breast cancer presentation. PATIENTS AND METHODS: All patients who underwent breast surgery from March 11, 2020 to May 30, 2020 were evaluated and considered as the Lockdown group. These patients were compared with similar patients of the previous year, the Pre-Lockdown group. RESULTS: A total of 432 patients were evaluated; n=223 and n=209 in the Lockdown and Pre-lockdown-groups, respectively. At univariate analysis, waiting times, lymph-nodes involvement and cancer grading, showed a statistically significant difference (p<0.05). Multivariate analysis identified waiting-time on list (OR=1.07) as a statistically significant predictive factor of lymph node involvement. CONCLUSION: Although we did not observe a clinically evident difference in breast cancer presentation, we reported an increase in lymph node involvement.

A protein corona sensor array detects breast and prostate cancers.

Following exposure to human plasma (HP), nanoparticles (NPs) are coated with a biomolecular layer referred to as a protein corona. We recently revealed that characterizing the protein coronas of various NPs may provide a unique opportunity for cancer identification and discrimination. In other words, protein corona profiles of several NPs, when being analyzed using classifiers, would provide a unique "fingerprint" for each type of disease. Here, we probed the capacity of the protein corona for the identification and discrimination of breast and prostate cancer patients from healthy individuals. Using three lipid NP formulations with distinct physical-chemical properties as a cross-reactive sensor array and a supervised random forest classifier, we identified a set of proteins that showed a significant difference in cancer patients and control subjects. Our data show that many of the corona proteins with the highest discrimination ability between oncological patients and healthy individuals are related to cellular and molecular aspects of breast and prostate cancers.

The Effect of Coronavirus (COVID-19) on Breast Cancer Teamwork: A Multicentric Survey.

BACKGROUND/AIM: Despite the large amount of clinical data available of Coronavirus-19 (COVID-19), not many studies have been conducted about the psychological toll on Health Care Workers (HCWs). PATIENTS AND METHODS: In this multicentric descriptive study, surveys were distributed among 4 different Breast Cancer Centers (BCC). BCCs were distinguished according to COVID-19 tertiary care hospital (COVID/No-COVID) and district prevalence (DP) (High vs. Low). DASS-21 score, PSS score and demographic data (age, sex, work) were evaluated. RESULTS: A total of 51 HCWs were analyzed in the study. Age, work and sex did not demonstrate statistically significant values. Statistically significant distribution was found between DASS-21-stress score and COVID/No-COVID (p=0.043). No difference was found in the remaining DASS-21 and PSS scores, dividing the HCWs according to COVID-19-hospital and DP. CONCLUSION: Working in a COVID-19-hospital represents a factor that negatively affects psychosocial well-being. However, DP seems not to affect the psychosocial well-being of BCC HCWs. During the outbreak, psychological support for low risk HCWs should be provided regardless DP.

Ultrasound-guided Interfascial Plane Blocks for Non-anesthesiologists in Breast Cancer Surgery: Functional Outcomes and Benefits.

AIM: Acute post-operative pain following modified radical mastectomy (MRM) in patients with breast cancer is challenging for anesthesiologists. This study aimed to prospectively compare the quality outcome of interfascial plane blocks performed with ultrasound guidance, and evaluate the consequences of sharing tasks with the breast surgeon. PATIENTS AND METHODS: The study involved 255 patients scheduled for unilateral MRM, who were divided into two groups: Pecs group: General anesthesia plus ultrasound-guided modified pectoral nerves blocks type I and II, including serratus and parasternal infiltration according to surgical requirements; and Control group: general anesthesia only. Quality was evaluated based on perioperative opioid consumption, reported pain intensity, rescue analgesic requirement, side-effects and length of hospital stay. Moreover, a breast surgeon with expertise in ultrasound-guided breast biopsy was trained to perform the blocks. The patient benefits from regional anesthesia delivered by a non-anesthesiologist were assessed. RESULTS: Significant reductions were noted in all of the following: Intraoperative opioid consumption (p<0.001), Numerating Rating Scale pain scores taken 0 and 24 h after surgery (p<0.001), post-operative analgesic administration (p<0.001), nausea and vomiting at 0, 6, and 12-h intervals (p<0.05), and hospital stay (p<0.001) were observed in the Pecs group compared with the control group. Furthermore, data obtained from patients receiving the block from the surgeon showed comparable benefits with no complications. CONCLUSION: Interfascial plane blocks may be an important alternative protocol in MRM, enhancing patient safety and cost benefits. Improvements in cross-disciplinary expertise through flexibility in the training of professionals with other backgrounds may provide effective analgesia and favorable outcomes.

Heterogeneous risk profiles among B3 breast lesions of uncertain malignant potential.

BACKGROUND: Most cases of breast lesions of uncertain malignant potential (B3) undergo surgical intervention. We aimed to analyze the outcome of B3 lesion subtypes in a large series of screen-detected cases. METHODS: We screened 2,986 core needle biopsies to classify B3 lesions. Positive predictive values (PPVs) for malignancy were calculated for a comprehensive risk characterization according to clinicopathologic and morphologic variables. RESULTS: B3 lesions comprised 35% atypical ductal hyperplasia (PPV = 20%), 16.7% flat epithelial atypia (PPV = 12%), 22.7% lobular neoplasia (PPV = 16.2%), 9% papillary lesion (PPV = 18.5%), 8.6% phyllodes tumor (PPV = 3.8%), and 8% radial scars (PPV = 4.1%) based on histopathologic diagnosis. Upgrade rates were 15.9% for calcifications, 13.7% for mass lesions, and 16.7% for architectural deformities, with 8.3% of malignant lesions classified as ductal carcinoma in situ and 6.7% as invasive cancers (PPV = 15%). CONCLUSION: B3 lesions entail a heterogeneous risk of malignancy, and careful radiologic-pathologic correlation is required for optimal treatment.

Direct-To-Implant and 2-Stage Breast Reconstruction After Nipple Sparing Mastectomy: Results of a Retrospective Comparison.

Breast reconstruction after nipple sparing mastectomy (NSM) plays, nowadays, a fundamental role in breast cancer management. There is no consensus on the best implant-based reconstruction technique, considering 2 stages (expander-prosthesis) or direct-to-implant (DTI). A retrospective review of consecutive adult female patients who underwent NSM with breast reconstruction over a 3-year period (January 2013 to December 2015) was performed. Patients were divided into 2 groups according to the type of reconstruction: expander/prosthesis (group A) and DTI (group B). Anamnestic data were collected. Number and type of procedures, complications and esthetic satisfaction were registered and compared. Fifty-six patients were included in group A (34.6%) and 106 in group B (65.4%). Complications associated with the 2 types of breast reconstruction were not different (P = 0.2). Patients in group A received a higher number of total surgical procedures (considering revisions, lipostructures and contralateral symmetrizations) than those in group B (2.5 ± 0.69 and 1.88 ± 1.02, P = 0.0001). Satisfaction with breast reconstruction resulted higher in group A (7.5 ± 2.6 and 6 ± 1.9, P = 0.0004). At the multivariate analysis, chemotherapy and radiotherapy were not correlated with complications, regardless of the group (odds ratio, 0.91 and 2.74, respectively). Radiotherapy and chemotherapy did not even influence the esthetic result, regardless of the group (P = 0.816 and P = 0.521, respectively). Prosthetic breast reconstructions, both in a single and in 2 stages, are welcomed by patients and have relatively low and almost equivalent complication rates, independent of other factors such as chemotherapy, radiotherapy, lymphadenectomy, smoking and age. In our experience, 2-stage breast reconstruction, although requiring more operations, is associated with a higher esthetic satisfaction. Patients who perform a DTI breast reconstruction after NSM should be informed of the high probability of surgical revision.

Limitations in predicting PAM50 intrinsic subtype and risk of relapse score with Ki67 in estrogen receptor-positive HER2-negative breast cancer.

PAM50/Prosigna gene expression-based assay identifies three categorical risk of relapse groups (ROR-low, ROR-intermediate and ROR-high) in post-menopausal patients with estrogen receptor estrogen receptor-positive (ER+)/ HER2-negative (HER2-) early breast cancer. Low risk patients might not need adjuvant chemotherapy since their risk of distant relapse at 10-years is below 10% with endocrine therapy only. In this study, 517 consecutive patients with ER+/HER2- and node-negative disease were evaluated for Ki67 and Prosigna. Most of Luminal A tumors (65.6%) and ROR-low tumors (70.9%) had low Ki67 values (0-10%); however, the percentage of patients with ROR-medium or ROR-high disease within the Ki67 0-10% group was 42.7% (with tumor sizes ≤2 cm) and 33.9% (with tumor sizes > 2 cm). Finally, we found that the optimal Ki67 cutoff for identifying Luminal A or ROR-low tumors was 14%. Ki67 as a surrogate biomarker in identifying Prosigna low-risk outcome patients or Luminal A disease in the clinical setting is unreliable. In the absence of a well-validated prognostic gene expression-based assay, the optimal Ki67 cutoff for identifying low-risk outcome patients or Luminal A disease remains at 14%.

Current trends and outcomes of breast reconstruction following nipple-sparing mastectomy: results from a national multicentric registry with 1006 cases over a 6-year period.

BACKGROUND: Reconstruction options following nipple-sparing mastectomy (NSM) are diverse and not yet investigated with level IA evidence. The analysis of surgical and oncological outcomes of NSM from the Italian National Registry shows its safety and wide acceptance both for prophylactic and therapeutic cases. A further in-depth analysis of the reconstructive approaches with their trend over time and their failures is the aim of this study. METHODS: Data extraction from the National Database was performed restricting cases to the 2009-2014 period. Different reconstruction procedures were analyzed in terms of their distribution over time and with respect to specific indications. A 1-year minimum follow-up was conducted to assess reconstructive unsuccessful events. Univariate and multivariate analyses were performed to investigate the causes of both prosthetic and autologous failures. RESULTS: 913 patients, for a total of 1006 procedures, are included in the analysis. A prosthetic only reconstruction is accomplished in 92.2 % of cases, while pure autologous tissues are employed in 4.2 % and a hybrid (prosthetic plus autologous) in 3.6 %. Direct-to-implant (DTI) reaches 48.7 % of all reconstructions in the year 2014. Prophylactic NSMs have a DTI reconstruction in 35.6 % of cases and an autologous tissue flap in 12.9 % of cases. Failures are 2.7 % overall: 0 % in pure autologous flaps and 9.1 % in hybrid cases. Significant risk factors for failures are diabetes and the previous radiation therapy on the operated breast. CONCLUSIONS: Reconstruction following NSM is mostly prosthetic in Italy, with DTI gaining large acceptance over time. Failures are low and occurring in diabetic and irradiated patients at the multivariate analysis.

Nipple sparing mastectomy: Surgical and oncological outcomes from a national multicentric registry with 913 patients (1006 cases) over a six year period.

BACKGROUND: Nipple sparing mastectomy is deemed surgically and oncologically safe based on a long lasting literature data from reviews of single institution series. This study aims at evaluating surgical and oncological outcomes of NSM on a large multi-institutional scale, by means of the Italian National registry. METHODS: In July 2011 a panel of Italian specialists agreed upon and designed a National database of NSM. Centers with at least 150 cancers per year and following the National follow-up schedule guidelines could participate inserting any NSM case performed, retrospectively and prospectively from that moment on. In March 2015 analysis of data was accomplished. Dataset for this study consists of cases performed in the period between January 1st 2009 and December 31st 2014. RESULTS: 913 Women were included in the analysis, for a total of 1006 procedures. Prophylactic mastectomies were 124 (12.3%). MRI utilization increased over time. NSM failure rate, with NAC removal for any reason was 11.5%. NAC necrosis rate was 4.8%. Larger skin-flap necrosis rate was 2.3%. Major surgical complications rate was 4.4%. Oncological outcomes were calculated among primitive EBC cases only: locoregional recurrences rate was 2.9%, NAC recurrence 0.7%. Systemic recurrence rate was 1.0%. Five deaths (0.7%) were registered. CONCLUSIONS: More than 10% of NSM procedures are prophylactic mastectomies. MRI is gaining more importance over time. Surgical and oncological results show that NSM is effective. This National multicentric analysis enables a comparison of results with no geographical differences and a "safe" state of the art of NSM in Italy.

In situ identification of CD44+/CD24- cancer cells in primary human breast carcinomas.

Breast cancer cells with the CD44+/CD24- phenotype have been reported to be tumourigenic due to their enhanced capacity for cancer development and their self-renewal potential. The identification of human tumourigenic breast cancer cells in surgical samples has recently received increased attention due to the implications for prognosis and treatment, although limitations exist in the interpretation of these studies. To better identify the CD44+/CD24- cells in routine surgical specimens, 56 primary breast carcinoma cases were analysed by immunofluorescence and confocal microscopy, and the results were compared using flow cytometry analysis to correlate the amount and distribution of the CD44+/CD24- population with clinicopathological features. Using these methods, we showed that the breast carcinoma cells displayed four distinct sub-populations based on the expression pattern of CD44 and CD24. The CD44+/CD24- cells were found in 91% of breast tumours and constituted an average of 6.12% (range, 0.11%-21.23%) of the tumour. A strong correlation was found between the percentage of CD44+/CD24- cells in primary tumours and distant metastasis development (p = 0.0001); in addition, there was an inverse significant association with ER and PGR status (p = 0.002 and p = 0.001, respectively). No relationship was evident with tumour size (T) and regional lymph node (N) status, differentiation grade, proliferative index or HER2 status. In a multivariate analysis, the percentage of CD44+/CD24- cancer cells was an independent factor related to metastasis development (p = 0.004). Our results indicate that confocal analysis of fluorescence-labelled breast cancer samples obtained at surgery is a reliable method to identify the CD44+/CD24- tumourigenic cell population, allowing for the stratification of breast cancer patients into two groups with substantially different relapse rates on the basis of CD44+/CD24- cell percentage.