Lymph Node Metastasis In Breast Cancer Research Articles

Artificial Intelligence-Based Breast Cancer Nodal Metastasis Detection: Insights Into the Black Box for Pathologists.

Nodal metastasis of a primary tumor influences therapy decisions for a variety of cancers. Histologic identification of tumor cells in lymph nodes can be laborious and error-prone, especially for small tumor foci. To evaluate the application and clinical implementation of a state-of-the-art deep learning-based artificial intelligence algorithm (LYmph Node Assistant or LYNA) for detection of metastatic breast cancer in sentinel lymph node biopsies. Whole slide images were obtained from hematoxylin-eosin-stained lymph nodes from 399 patients (publicly available Camelyon16 challenge dataset). LYNA was developed by using 270 slides and evaluated on the remaining 129 slides. We compared the findings to those obtained from an independent laboratory (108 slides from 20 patients/86 blocks) using a different scanner to measure reproducibility. LYNA achieved a slide-level area under the receiver operating characteristic (AUC) of 99% and a tumor-level sensitivity of 91% at 1 false positive per patient on the Camelyon16 evaluation dataset. We also identified 2 "normal" slides that contained micrometastases. When applied to our second dataset, LYNA achieved an AUC of 99.6%. LYNA was not affected by common histology artifacts such as overfixation, poor staining, and air bubbles. Artificial intelligence algorithms can exhaustively evaluate every tissue patch on a slide, achieving higher tumor-level sensitivity than, and comparable slide-level performance to, pathologists. These techniques may improve the pathologist's productivity and reduce the number of false negatives associated with morphologic detection of tumor cells. We provide a framework to aid practicing pathologists in assessing such algorithms for adoption into their workflow (akin to how a pathologist assesses immunohistochemistry results).

Regulation of RAB22A by mir-193b inhibits breast cancer growth and metastasis mediated by exosomes.

Breast cancer is one of the main types of cancer affecting the health of females worldwide. Despite improvements in therapeutic approaches, cancer patients succumb to the disease due to metastasis itself, rather than the primary tumor from which metastases arise, emphasizing the need for the better understanding of the biological bases that contribute to disease progression. RAB22A, a member of the proto-oncogene RAS family, plays an important role in the formation, trafficking and metabolism of exosomes, and is associated with the occurrence and development of multiple human cancers. In this study, we demonstrate that the upregulation of RAB22A is associated with breast cancer progression and lymph node metastasis. We identified a signature of RAB22A and miR-193b that exhibited a negative association in metastatic as opposed to the surrounding normal cells, and RAB22A was identified as the target gene of miR-193b. While RAB22A was found to regulate exosomes-mediated breast cancer cell proliferation, invasion and migration, these biological characteristics were diminished in the breast cancer cells in which the RAB22A gene was knocked down or in the cells in which the exosomes were dissolved by proteinaseK/RNase treatment. On the whole, the findings of this study demonstrate the critical role that miR-193b plays in the regulation of RAB22A-mediated exosome function during cancer growth and metastasis, which may have significant implications on cancer therapy.

Internal mammary lymph node metastases in breast cancer: what should radiologists know?

The internal mammary lymph node (IMLN) chain is a pathway through which breast lymphatic drainage flows. The internal mammary lymphatic vessel runs around the internal mammary artery and veins with IMLN in the parasternal intercostal spaces. IMLN metastasis, which forms a part of clinical TNM staging, may negatively affect the prognosis of primary breast cancer patients. IMLN metastasis is clinically detected using ultrasound, computed tomography, magnetic resonance imaging, and 18F-deoxyglucose positron emission tomography computed tomography. The uptake of radioactive tracers in IMLN with clinically negative axillary lymph nodes is often identified using sentinel lymph node mapping (SLNM) in primary breast cancer patients. The indication for IMLN biopsy or resection that is clinically detected or visualized using SLNM is controversial. The clinically suspicious IMLN may be considered for ultrasound-guided fine-needle aspiration. First IMLN recurrence needs to be biopsied. Irradiation of the breast, chest wall, and/or regional nodal irradiation, including IMLN, following lumpectomy or postmastectomy is recommended. Although radiation therapy for IMLN recurrence may improve clinical outcomes, it is also associated with pulmonary and cardiac toxicities. This review covers the local anatomy of IMLN, lymph drainage and image findings of IMLN with a discussion.

ZNF259 promotes breast cancer cells invasion and migration via ERK/GSK3β/snail signaling.

PurposeZinc finger protein 259 (ZNF259), also known as ZPR1, is a zinc finger-containing protein that can bind the intracellular tyrosine kinase domain of EGFR. At present, our knowledge on ZNF259 in cancers is limited. Here, we aimed to explore the biological functions of ZNF259 in breast cancer and reveal their mechanisms.Patients and methodsThe expression of ZNF259 was measured in 133 cases of breast cancer by immunohistochemistry. The online database Kaplan–Meier (KM) Plotter Online Tool was used to analyze the relationship between ZNF259 expression and breast cancer patient survival prognosis. Plasmid transfection and small interfering RNA and inhibitor treatments were carried out to explore the functions of ZNF259 in breast cancer cell lines and its potential mechanism. Matrigel invasion and wound healing assays were performed to detect the invasion and migration ability of cancer cells. In addition, protein expressions in tissues and cells were determined by Western blotting.ResultsZNF259 expression was much higher in breast cancer cells than in the adjacent normal breast duct glandular epithelial cells (75.94% vs 7.52%, P<0.001) and was closely related to the breast cancer patients’ TNM stages (P=0.013) and lymph node metastasis (P=0.021). Knockdown of ZNF259 could downregulate p-ERK, p-GSK3β, and Snail expression, and upregulate the expression of E-cadherin and ZO-1, and then it also inhibited invasion and migration by the breast cancer cell lines MCF-7 and MDA-MB-231. Correspondingly, ZNF259 transfection could upregulate p-ERK, p-GSK3β, and Snail expression, and downregulate E-cadherin and ZO-1 expression, which led to stronger invasion and migration abilities of cancer cells. Furthermore, the ERK inhibitor U0126 could reverse all these effects induced by ZNF259 transfection.ConclusionZNF259 could promote breast cancer cell invasion and migration by activating the ERK/GSK3β/Snail signaling pathway.

CXCL1 derived from tumor-associated macrophages promotes breast cancer metastasis via activating NF-\u03baB/SOX4 signaling

Tumor-associated macrophages (TAMs) have been implicated in the promotion of breast cancer growth and metastasis, and multiple TAM-secreted cytokines have been identified associating with poor clinical outcomes. However, the therapeutic targets existing in the loop between TAMs and cancer cells are still required for further investigation. Here in, cytokine array validated that C-X-C motif chemokine ligand 1 (CXCL1) is the most abundant chemokine secreted by TAMs, and CXCL1 can promote breast cancer migration and invasion ability, as well as epithelial–mesenchymal transition in both mouse and human breast cancer cells. QPCR screening further validated SOX4 as the highest responsive gene following CXCL1 administration. Mechanistic study revealed that CXCL1 binds to SOX4 promoter and activates its transcription via NF-κB pathway. In vivo breast cancer xenografts demonstrated that CXCL1 silencing in TAMs results in a significant reduction in breast cancer growth and metastatic burden. Bioinformatic analysis and clinical investigation finally suggested that high CXCL1 expression is significantly correlated with breast cancer lymph node metastasis, poor overall survival and basal-like subtype. Taken together, our results indicated that TAMs/CXCL1 promotes breast cancer metastasis via NF-κB/SOX4 activation, and CXCL1-based therapy might become a novel strategy for breast cancer metastasis prevention.

Elastic scattering spectroscopy for early detection of breast cancer: partially supervised Bayesian image classification of scanned sentinel lymph nodes.

Sentinel lymph node biopsy is a standard diagnosis procedure to determine whether breast cancer has spread to the lymph glands in the armpit (the axillary nodes). The metastatic status of the sentinel node (the first node in the axillary chain that drains the affected breast) is the determining factor in surgery between conservative lumpectomy and more radical mastectomy including axillary node excision. The traditional assessment of the node requires sample preparation and pathologist interpretation. An automated elastic scattering spectroscopy (ESS) scanning device was constructed to take measurements from the entire cut surface of the excised sentinel node and to produce ESS images for cancer diagnosis. Here, we report on a partially supervised image classification scheme employing a Bayesian multivariate, finite mixture model with a Markov random field (MRF) spatial prior. A reduced dimensional space was applied to represent the scanning data of the node by a statistical image, in which normal, metastatic, and nonnodal-tissue pixels are identified. Our results show that our model enables rapid imaging of lymph nodes. It can be used to recognize nonnodal areas automatically at the same time as diagnosing sentinel node metastases with sensitivity and specificity of 85% and 94%, respectively. ESS images can help surgeons by providing a reliable and rapid intraoperative determination of sentinel nodal metastases in breast cancer.

Abstract 1104: Tumor-associated macrophages-secreted CXCL1 promotes breast cancer metastasis via activating NF-κB/SOX4 signaling

Abstract Purpose: Secreted cytokines play a significant role in modulating cancer metastatic microenvironment, especially for tumor associated macrophages (TAMs). The current study aimed to determine the significance of TAMs-secreted CXCL1 in regulating cancer metastasis and its underlying mechanisms. Experimental Design: Cytokine array was utilized to screen the highest chemokine secreted from TAMs. In vivo immunohistochemistry assay and in vitro wound healing or transwell experiments were carried out to demonstrate the metastatic-promoting effects of TAMs/CXCL1. QPCR profile screening was used to identify CXCL1 targeting metastasis-related gene. In vivo luciferase imaging experiments were designed to confirm the therapeutic significance of CXCL1 silencing on breast cancer metastasis. The clinical significance of CXCL1 was analyzed by human tissue microarray and bioinformatic analysis. Results: CXCL1 was validated as the highest secreted protein released from TAMs, and TAMs-derived CXCL1 could promote breast cancer migration and invasion ability, as well as epithelial-mesenchymal transition (EMT) process. QPCR screening validated SOX4 as the most responsive gene following CXCL1 administration. Mechanistic study revealed that CXCL1 activates SOX4 transcription via NF-κB pathway. CXCL1 silencing in TAMs resulted in a significant reduction in breast cancer growth and metastatic burden. Clinical investigation suggests that high CXCL1 expression was significantly correlated with breast cancer lymph node metastasis, poor overall survival and basal-like subtype. Conclusions: TAMs/CXCL1 promotes breast cancer metastasis via NF-κB/sox4 activation, and CAV1 based therapy might become a novel strategy for breast cancer metastasis prevention. Citation Format: Zhiyu Wang, Neng Wang, Weiping Liu, Yifeng Zheng, Shengqi Wang. Tumor-associated macrophages-secreted CXCL1 promotes breast cancer metastasis via activating NF-κB/SOX4 signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 1104.

Inhibition of the AnxA1/FPR1 autocrine axis reduces MDA-MB-231 breast cancer cell growth and aggressiveness in vitro and in vivo

Breast Cancer (BC) is a highly heterogeneous disease whose most aggressive behavior is displayed by triple-negative breast cancer (TNBC), which lacks an efficient targeted therapy. Despite its controversial role, one of the proteins that having been linked with BC is Annexin A1 (AnxA1), which is a Ca+2 binding protein that acts modulating the immune system, cell membrane organization and vesicular trafficking. In this work we analyzed tissue microarrays of BC samples and observed a higher expression of AnxA1 in TNBCs and in lymph node metastasis. We also observed a positive correlation in primary tumors between expression levels of AnxA1 and its receptor, FPR1. Despite displaying a lesser strength, this correlation also exists in BC lymph node metastasis. In agreement, we have found that AnxA1 was highly expressed and secreted in the TNBC cell line MDA-MB-231 that also expressed high levels of FPR1. Furthermore, we demonstrated, by using the specific FPR1 inhibitor Cyclosporin H (CsH) and the immunosuppressive drug Cyclosporin A (CsA), the existence of an autocrine signaling of AnxA1 through the FPR1. Such signaling, elicited by AnxA1 upon its secretion, increased the aggressiveness and survival of MDA-MB-231 cells. In this manner, we demonstrated that CsA works very efficiently as an FPR1 inhibitor. Finally, by using CsA, we demonstrated that FPR1 inhibition decreased MDA-MB-231 tumor growth and metastasis formation in nude mice. These results indicate that FPR1 inhibition could be a potential intervention strategy to manage TNBCs displaying the characteristics of MDA-MB-231 cells. FPR1 inhibition can be efficiently achieved by CsA.

Extent of breast cancer type 1 promoter methylation correlates with clinicopathological features in breast cancers.

The current meta-analysis investigated the correlation between breast cancer type 1 (BRCA1) promoter methylation and the clinicopathological features of breast cancer (BC). An electronic literature search was performed to identify and select cohort studies, by employing stringent inclusion and exclusion criteria, for data relevant to promoter methylation of BRCA1 and BC. Statistical analysis of the extracted data was performed using comprehensive meta-analysis 2.0 software (CMA 2.0) (Biostat Inc., Englewood, New Jersey, USA). A total of 125 published studies were retrieved from the literature search, and finally, 18 cohort studies meeting our inclusion criteria were incorporated into our meta-analysis. The 18 studies contained a total of 3213 BC patients. Meta-analysis results revealed that BRCA1 promoter methylation in BC patients with high and moderately differentiated tumors (I-II) was significantly lower than patients with poorly-differentiation tumors (III) (odds ratio [OR] =0.450, 95% confidence interval [95% CI] =0.241-0.838, P = 0.012). BRCA1 promoter methylation in BC patients with lymph node (LN) metastasis was significantly higher than patients without LN metastasis (OR = 2.244, 95% CI = 1.278-3.940, P = 0.005). The results of ethnicity-based subgroup analysis showed a significant difference in histological grade of BC on Asians, LN metastasis of BC in Asians and Caucasians, subtypes of BC in Caucasians, and age at diagnosis of BC patients in Caucasians (all P < 0.05). Our meta-analysis revealed that BRCA1 promoter methylation status is linked to tumor grade and LN metastasis of BC.

Stromal immunoglobulin κC expression is associated with initiation of breast cancer in TA2 mice and human breast cancer.

The initiation of spontaneous breast cancer (SBC) in Tientsin Albino 2 (TA2) mice is related to mouse mammary tumor virus (MMTV) infection, and MMTV amplification is hormonally regulated. To explore the insertion site of MMTVLTR in TA2 mouse genome, reverse PCR and nested PCR were used to amplify the unknown sequence on both sides of the MMTV‐LTRSAG gene in SBC and normal breast tissue of TA2 mice. Furthermore, the clinicopathological significance of the insertion site was evaluated in 43 samples of normal breast tissue, 46 samples of breast cystic hyperplasia, 54 samples of ductal carcinoma in situ, 142 samples of primary breast cancer and 47 samples of lymph node metastatic breast cancer by RNA in situ hybridization. We confirmed that the insertion site of the MMTV‐LTRSAG gene was located between Igκv2‐112 and Igκv14‐111 in chromosome 6 of TA2 mouse. IG κC was localized in the stromal cells of TA2 mouse with SBC and in human breast cancer tissues. Tumor cells were negative for IG κC in RNA in situ hybridization. The positive staining index of IG κC in stromal cells was the highest in lymph node metastatic breast cancer, followed by primary breast cancer, ductal carcinoma in situ, and breast cystic hyperplasia. Furthermore, the positive staining index of IG κC was related to the expression of ER, PR, HER2 and Ki‐67. Our findings showed that stromal IG κC expression was associated with the initiation of SBC in TA2 mice. IG κC may be a high‐risk factor for the initiation and progression of human breast cancer.

Expression of Lymphangiogenesis Markers and Correlation with Lymph Nodal Metastasis in Breast cancer

AimTo study expression of VEGF and D II40 in breast cancer and correlate with lymph nodal metastasis.Materials &amp; MethodsFormalin fixed paraffin embedded blocks of breast cancer were retrieved and 4 mu thick sections obtained&amp; stained with routine ‘H&amp;E’. Sections were cut on APES (3‐Triethoxysilylpropylamine) coated slides &amp; immunohistochemistry (IHC) was performed using Streptavidin Biotin Peroxidase technique using monoclonal antibodies against VEGF and DII 40 on coated sections of cases &amp; controls. IHC stains were reviewed in conjunction with H&amp;E. Intensity of VEGF expression &amp; its distribution in the tumour were observed under high power magnification &amp; semi‐quantitatively analyzed. Brown granules in the cytoplasm/membrane of the cells were considered as +ve for VEGF expression when the proportion of immunoreactive cells was ≥5%. ; 5–25% staining, weakly +ve (+) 26–50% staining,+ve (++); &gt;50% staining, strongly +ve (+++). For statistical analysis, samples with 0/+ staining considered in low expression group, while ++/+++ expression were considered as high expression. For DII40 well demarcated vessels showing positivity in endothelial cells were counted per 10 hpf after selecting hot spots at scanner view. For statistical analysis, 0 was considered as no expression and 1/10HPF was considered as high expression.Results48 tissue specimens were included in the study. High VEGF expression (Fig 1) was seen 33/43 patients (76.74%%; 95% CI 61.37%–88.24%) and high DII 40 expression (Fig 2) in 30/44 (68.18%; 95% CI 52.42%–81.39%). Variation in expression of VEGF and DII 40 in relation to each other is given in figure 1. Of the 33 patients with high VEGF expression, DII 40 was expressed in 26 of them (78.79%). There is a strong association between VEGF expression and DII 40 expression (p&lt;0.01). The scatter diagram of VEGF and DII 40 expression showed a significant trend (Figure 3 ) indicating that as VEGF expression increases so does the DII 40 expression. The variance explained by VEGF level in the variance of DII 40 as measured by the correlation (R2) was 0.1265 (p=0.02). When VEGF expression was compared with LN status, low expression of VEGF was found in 38.47 % (5/13) of SLN positive and 16.67 % (5/30) of negative senitnel node patients. High expression of VEGF was in 61.53% of positive patients as compared to 83.33% of negative sentinel node patients. Results of VEGF expression and sentinel node status revealed no statistically significant relation between expression of VEGF and LN involvement. No D II 40 expression of was found in 38.46% (5/13) % of LN +ve patients as compared to 29.03% (9/31) in node negative. High D II 40 expression was found 61.54% LN+ve patients and 70.97% of snode negative. Results of DII 40 expression and LN status revealed no statistically significant relation between expression of DII 40 and node involvement. When the expression of VEGF and D II 40 was compared with tumour characteristics there was no statistically significant relation with tumour size, tumour type, grade, ER/PR staus and Her2neu status.ConclusionsLympahngiogenesis marker (VEGF) expression is high in most tumours and there is significant association between lymph vessels detection as studied by D II 40 expression and lymphangiogenesis marker expression as studied VEGF expresssion. However, no association was found between expression of VEGF and D II 40 with neither sentinel / non‐sentinel nodal status nor any of the primary tumour characteristics.Support or Funding InformationnoneThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Detecting the role of CCR7-CCL21/CCL19 axis in Breast Cancer Progression and lymph node Metastasis Incidence

Background: Metastasis is the main cause of death in breast cancer patients. Lymph node metastasis is associated with poor overall survival and is often associated with recurrences. The molecular mechanisms controlling tumor metastasis are complex and there is an emerging evidence supporting the direct involvement of the chemokines and their receptors in this process. To the interest of this study, previous data from the literature show that the CCL21 and CCL19 and their receptor CCR7 are expressed in breast cancer and may be associated with lymph node metastasis. The aim of this study is to assess the expression of CCL19/CCL21 and their receptor CCR7 in breast cancer tissues and both negative and positive regional lymph nodes using RT-PCR technology . Material and Methods: Formalin fixed paraffin embedded breast cancer specimens obtained from mastectomy were included in the study. RNA isolation using Acidic Phenol method was performed, and then RT-PCR was conducted on both primary tumor and corresponding lymph nodes using specific primers for the studied CCL21/CCL19 chemokines, CCR7 receptor and the positive control GAPDH. Results: The results of this study have revealed a significant positive correlation in mRNA expression in the tumor samples between CCL21 vs.CCL19 (R2=0.9,P-value= 0.02), as well as between CCR7 vs CCL21, and CCR7vs.CCL19 (R2=0.87, P value= 0.04) (R2=0.98,P value= 0.002) respectively. Moreover, there was a significant positive correlation (R2=0.9 , P value= 0.02) in CCL19 expression in primary and secondary tumor. Furthermore, the expression of CCL21 chemokine in negative lymph nodes was more than the expression of it in positive lymph nodes with significant importance (P value = 0.01). The Study of the correlation between CCL21/CCL19 expression in breast tumors and the clinical and pathological parameters, has illustrated a significant correlation between CCL21 vs. CCL19 mRNA expression in breast tumors and triple negative of PR, ER, Her-2 receptors. Conclusion: The results of the study have revealed a potential role of CCR7-CCL21/CCL19 axis in breast cancer. Moreover the results suggest an important role of CCL19 in Lymph nodes metastasis compared to CCL21. Furthermore, the study has showed that CCL21/CCL19 chemokines are expressed in breast tumors with poor prognosis.

Use of Sentinel Lymph Node Biopsy after Neoadjuvant Chemotherapy in Patients with Axillary Node-Positive Breast Cancer in Diagnosis.

PurposeThis study aimed to evaluate the effects of sentinel lymph node biopsy (SLNB) on recurrence and survival after neoadjuvant chemotherapy (NAC) in breast cancer patients with cytology-proven axillary node metastasis.MethodsWe selected patients who were diagnosed with invasive breast cancer and axillary lymph node metastasis and were treated with NAC followed by curative surgery between January 2007 and December 2014. We classified patients into three groups: group A, negative sentinel lymph node (SLN) status and no further dissection; group B, negative SLN status with backup axillary lymph node dissection (ALND); and group C, no residual axillary metastasis on pathology with standard ALND.ResultsThe median follow-up time was 51 months (range, 3–122 months) and the median number of retrieved SLNs was 5 (range, 2–9). The SLN identification rate was 98.3% (234/238 patients), and the false negative rate of SLNB after NAC was 7.5%. There was no significant difference in axillary recurrence-free survival (p=0.118), disease-free survival (DFS; p=0.578) or overall survival (OS; p=0.149) among groups A, B, and C. In the subgroup analysis of breast pathologic complete response (pCR) status, there was no significant difference in DFS (p=0.271, p=0.892) or OS (p=0.207, p=0.300) in the breast pCR and non-pCR patients.ConclusionThese results suggest that SLNB can be feasible and oncologically safe after NAC for cytology-determined axillary node metastasis patients and could help reduce arm morbidity and lymphedema by avoiding ALND in SLN-negative patients.

Ultrasound and Elastographic Features predicting Axillary Nodal Metastasis in Breast Cancer

AbstractLymph node metastasis is an important prognostic factor in cases of carcinoma breast. In this pictorial essay, various sonographic and elastographic signs of lymph node metastasis are described with classical images.

Correlation between Ultrasound Appearance of Small Breast Cancer and Axillary Lymph Node Metastasis

To study the correlation of ultrasonographic signs of small breast cancer (maximum diameter ≤2.0 cm) with axillary lymph node metastasis, pre-operative ultrasonographic images of 153 small breast malignant neoplasms in 143 breast cancer patients were analyzed according to their pathologic features. Of the small breast tumors included, 47 showed axillary lymph node metastasis. Diagnosis of all patients was obtained with radical axillary surgery or a sentinel lymph node biopsy procedure. Ultrasonographic signs included irregular shape, microlobulated contour, spiculation, microcalcification, posterior echo attenuation, blood-flow grade, perforating vessels, changes in fascia or cooper's ligament and maximum cortical thickness of lymph nodes. The relationship between ultrasonographic features and axillary lymph node metastasis was analyzed using a chi-square test for univariate distributions and logistic regression for multivariate analysis. A logistic regression model was established by taking the pathologic diagnosis of lymph node metastasis as the dependent variable and the ultrasonographic signs of each small breast cancer as independent variables. In small breast cancer, characteristics such as perforating vessels and maximum cortical thickness of lymph nodes >3.0 mm correlated well with axillary lymph node metastasis as determined by univariate analysis (χ2 = 13.945, 51.276, respectively, p < 0.05) and multivariate analysis (OR = 48.783, 46.754, respectively, p < 0.05).

Validation of a Novel Diagnostic Kit Using the Semidry Dot-Blot Method to Detect Metastatic Lymph Nodes in Breast Cancer: Distinguishing Macrometastases From Nonmacrometastases

BackgroundThe semidry dot-blot method is a diagnostic procedure for detecting lymph node (LN) metastases using the presence of cytokeratin (CK) in lavage fluid from sectioned LNs. We evaluated 2 novel kits that use newly developed anti-CK-19 antibodies to diagnose LN metastases in breast cancer. Patients and MethodsWe examined 159 LNs dissected that we sliced at 2-mm intervals and washed with phosphate-buffered saline. The suspended cells in the lavage were centrifuged and lysed to extract protein. This extracted protein was used with a low-power and a high-power kit to diagnose LN metastasis. Diagnoses on the basis of the kits were compared with pathological diagnoses. ResultsOf the 159 LNs, 68 were assessed as positive and 91 as negative in permanent section examination. Sensitivity, specificity, and accuracy of the low-power kit for detecting LN metastases was 83.8%, 100%, and 93.1%, respectively. Those of the high-power kit were 92.6%, 92.3%, and 92.5%, respectively. Combining the low- and high-power kit results, those for distinguishing macrometastases were 94.5%, 95.2%, and 95.0%, respectively. Diagnosis was achieved in approximately 20 minutes, at a cost of less than $30 USD. ConclusionThe kits were accurate, fast, and cost-effective in diagnosing LN metastases without the loss of LN tissue.

The correlation study between greyscale ultrasonography of invasive breast cancer and axillary lymph node involvement

Objective To investigate the relationship of greyscale ultrasonographic signs and clinical pathological characteristics with axillary lymph node metastasis in invasive breast cancer. Methods Three hundred and thirty-five patients with pathologically confirmed breast cancer were retrospectively analyzed, which categorized into 2 groups, including lymph node metastasis group and lymph node non-metastasis group. The clinical and pathological characteristics included age, primary tumor size, histological grade, and molecular subtypes. The ultrasonographic features were examined including orientation, shape, margin, echo pattern, posterior acoustic feature, and calcification. Chi-Square test and Logistic regression analysis were performed to analyze the relationship of clinical pathological characteristics and greyscale ultrasonographic signs with axillary lymph node metastasis. Results Age ( 10) contributed to axillary lymph node metastasis in 335 cases, according to univariate analysis (P<0.05). Multivariate Logistic regression analysis showed age (OR=0.573, P=0.019), the primary tumor size (OR=2.359, P=0.001), histological grade (OR=0.529, P=0.008), and number of margin angulation (OR=1.889, P=0.031) were risk factors of axillary lymph node metastasis in breast cancer. Conclusions Univariate and multivariate analyses show the correlation between ultrasonographic features and axillary lymph node involvement. Combining with clinical pathological data, it can provide a predictor of axillary lymph node metastasis. Key words: Ultrasonography; Invasive breast cancer; Lymph node metastasis; Clinical pathology

Correlation of expression of epithelial cell adhesion molecule and lymph node metastasis of breast cancer

Objective To investigate the expression and significance of epithelial cell adhesion molecule (Ep-CAM) in breast cancer. Methods Twenty-three cases of breast cancer tissue samples and paired lymph node metastases confirmed pathologically were collected. Isobaric tags for relative and absolute quantitation (iTRAQ) proteomics technology was used to screen and identify the differentially expressed proteins between primary tumor and lymph node metastasis of breast cancer. The expression of Ep-CAM was detected by Western blotting in 4 cases of primary breast cancer tissues and paired lymph node metastases. And the expressions of Ep-CAM in 252 cases of breast lesions were detected by immunohistochemical method. Results Quantitative proteomic examination results showed that differentially expressed proteins existed in breast cancer primary tumor and lymph node metastasis, and the expression of Ep-CAM in metastatic lesions was higher than that in primary tumor. Western blotting results showed that the expression of Ep-CAM in metastatic lesions (1.46±0.22) was higher than that in primary tumor (1.16±0.09), which was consistent with the results of proteomic. The immunohistochemical results showed that the positive expression rate of Ep-CAM in lymph node metastasis tissues (93.16%, 109/117) was significantly higher than that in primary breast cancer without metastasis (72.73%, 64/88), and the difference was statistically significant (χ2=15.921, P=0.000). The positive expression rate of Ep-CAM in primary breast cancer with lymphatic metastasis (72.65%, 85/117) was lower than that in paired lymph node metastases (P=0.001). Conclusion Ep-CAM is diffe-rentially expressed in primary tumor and lymph node metastasis of breast cancer, which may be related to the lymphatic metastasis of breast cancer. Key words: Cell adhesion molecule; Breast neoplasms; Proteomics; Immunohistochenmistry

P29 - An external quality control to validate One Step Nucleic Acid (OSNA) amplification accuracy for detection of breast cancer lymph node metastasis

P29 - An external quality control to validate One Step Nucleic Acid (OSNA) amplification accuracy for detection of breast cancer lymph node metastasis

P198 - MRI-based predictive factors of axillary lymph node metastases in breast cancer

P198 - MRI-based predictive factors of axillary lymph node metastases in breast cancer