Lymphatic Metastasis Of Breast Cancer Research Articles

Diagnosis of Lymphatic Metastasis in Breast Cancer Using Nanoparticle Technology - Diagnosis, Therapy, Imaging, Treatment

Breast cancer is a major cause of patient death rates, frequently leading to life-changing repercussions even after survival is attained. This paper aims to investigate therapeutic alternatives employing nanoparticles to specifically target and treat lymphatic metastasis, which is a highly dangerous characteristic of breast cancer. This work explores the effectiveness and importance of using nanoparticle-based therapeutics to prevent the harmful consequences of breast cancer progression. The paper begins by discussing the progress of lymphatic metastasis and then delves into the use of nanoparticle technology in imaging techniques, diagnostic methods, and therapy tactics. This section provides detailed information on primary targeting treatments, including chemotherapy specifically targeting cancer stem cells, induction of tumour cell death, suppression of Epithelial-Mesenchymal Transition (EMT), manipulation of the Tumour Microenvironment (TME), and improvement of the immune response. In addition, the research explores the use of nanoparticle technology in treatment plans, specifically focusing on its super magnetic capabilities and the application of gold nanoparticles, nanodiamonds, and other related qualities. Nanoparticle technology presents an optimistic strategy to address lymphatic metastasis in breast cancer. Nanoparticles can be used to deliver drugs or therapeutic substances directly to cancerous tumours, specifically targeting cancer cells to either destroy them or slow their growth. This strategy provides a solution for the administration of pharmaceuticals or substances that may provide challenges when delivered using conventional methods. Furthermore, nanoparticles facilitate the visualisation of tumours, aiding healthcare professionals in evaluating the severity of malignancy and formulating suitable treatment strategies. A comprehensive discussion has been conducted on several nanoparticles employed for inhibiting the dissemination of cancer cells from the primary organ to secondary organs. After successfully overcoming breast cancer, patients remain susceptible to post-surgical metastases in vital organs such as the lungs, brain, and bones. The advancements achieved through nanoparticle technology are highly significant. The discussion has focused on experimental evidence offered by researchers who mostly conducted studies on mice to support their findings.

A retrospective study on improving the accuracy of radiotherapy for patients with breast cancer with lymph node metastasis using Styrofoam.

To retrospectively analyze the accuracy of radiotherapy using cone beam computed tomography (CBCT), Styrofoam fixation, and breast bracket fixation in the chest wall target area and supraclavicular lymphatic drainage area (supraclavicular target area) of patients with breast cancer.and compare the setting efficiency and comfort satisfaction. A total of 65 patients with postoperative lymphatic metastasis of breast cancer, including 36 cases of Styrofoam fixation and 29 cases of breast bracket fixation, were recruited from March 2021 to August 2022 and retrospectively analyzed. All the patients underwent CBCT scans weekly, and the setup errors of the chest wall and supraclavicular target volume were compared and recorded. The planning target volume (PTV) margins of the two groups were calculated using the correlation MPTV = 2.5Σ + 0.7σ. The setup time and comfort satisfaction scores of the two groups were recorded and analyzed. The correlations among errors in each direction were analyzed using the Pearson correlation analysis. There was a significant difference in the left-right direction (X) axis of the chest wall target area between the Styrofoam and breast bracket groups (1.59 ± 1.47 mm vs. 2.05 ± 1.64 mm, P = 0.012). There were statistical differences in the ventrodorsal direction (Z) and bed angle of the supraclavicular target area, the data were (1.36 ± 1.27 mm vs. 1.75 ± 1.55 mm, P = 0.046; 0.47 ± 0.47° vs. 0.66 ± 0.59°, P = 0.006, respectively). In the X, Y, and Z directions, the respective PTV margins of the two groups in the chest wall target area were 5.01 mm, 5.99 mm, and 5.47 mm in the Styrofoam group, while those in the breast bracket group were 6.10 mm, 6.34 mm, and 6.10 mm, respectively. Moreover, the PTV margins of the supraclavicular target in the three directions were 3.69 mm, 3.86 mm, and 4.28 mm in the Styrofoam group, while those in the breast bracket group were 3.99 mm, 3.72 mm, and 5.45 mm, respectively. The setup time of the two groups was 3.4 ± 1.1 min and 5.5 ± 3.1 min (P = 0.007). The subjective comfort satisfaction scores of the two groups were 27.50 ± 1.24 and 25.44 ± 1.23 (P < 0.001). The application of Styrofoam fixation in radiotherapy of breast cancer in the supraclavicular lymph node area has several advantages as compared to breast bracket fixation, including higher positioning accuracy, smaller external expansion boundary, improved work efficiency, and patients' comfort, which might provide a reference for clinical work.

Lymphatic Architecture of Sappey Plexus and Lymphatic Metastasis of Breast Cancer

Согласно статистике, количество женщин с диагностированным раком молочной железы в мире на 2020 г. составило 2,3 млн чел. Количество смертей от рака молочной железы составило примерно 690 000 (6,9 %) случаев. Около 70 % умерших женщин имели метастазы. Одним из основных способов метастазирования рака молочной железы считается лимфогенное распространение, что делает крайне необходимым понимание топографо-анатомических особенностей лимфатического русла молочной железы, что может стать ключом к пониманию путей метастазирования, позволит более точно определить объем оперативного вмешательства и прогнозировать вероятность возникновения отдаленных метастазов. В рамках данного литературного обзора подробно разобраны исторические аспекты изучения анатомии глубоких и поверхностных лимфатических сосудов молочной железы, рассмотрены особенности их топографии по данным современных исследований и представлены различные варианты лимфогенного метастазирования на примере клинических случаев. Обсуждаются некоторые результаты исследования, проведенного авторами данной статьи, в котором методами цветной лимфографии, классической гистологии и иммуногистохимии с использованием антител к подопланину (D2-40), гладкомышечному актину (SMA), панцитокератину AE1/AE3 и транскрипционному фактору GATA3, изучено на макро- и микроскопическом уровне подареолярное лимфатическое сплетение Саппея, показана его связь с лимфатической системой молочной железы и выявлены признаки, подтверждающие участие подареолярного лимфатического сплетения в процессе лимфогенного метастазирования при раке молочной железы с метастазами в подмышечных лимфатических узлах. Для подготовки обзора проведен поиск литературы по базам данных Scopus, Web of Science, Medline, PubMed, CyberLeninka, РИНЦ и CNKI. При анализе использованы источники, индексируемые в базах данных Scopus и Web of Science (78 %), РИНЦ и CNKI (22 %); 35 % работ опубликовано за последние 5 лет. Использовано 37 источников для написания данного литературного обзора.

Does VEGF Expression Correlate with Lymphatic Metastasis in Breast Cancer?

IntroductionPrimary tumor characteristics are increasingly used for prognostication, predicting lymph node metastasis and planning adjuvant treatment in breast cancer. Vascular Endothelial Growth factor (VEGF) has been postulated to stimulate lymph vessels development and growth in tumors. Few studies on cell lines have documented how the new lymphatics develop or tumor cells invade these lymphatics. However, in human breast cancer the literature is sparse. Hence, this study is done to see the expression of VEGF and its correlation with lymphatic invasion in breast cancer patients.Materials &amp; MethodsFormalin fixed paraffin embedded blocks of breast cancer specimens and controls were retrieved &amp; 4u thick sections stained with H&amp;E stains for morphological evaluation. Serial sections were cut on APES (3‐Triethoxysilylpropylamine) coated slides for performing immunohistochemistry using monoclonal antibodies against VEGF and DII40. The expression intensity of VEGF and its distribution in the tumor samples were observed and semi‐quantitatively analyzed. VEGF expression was interpreted as follows: negative (0): &lt;5% staining; weak (+): 5–25% staining; positive (++): 26–50% staining; strongly positive (+++): &gt;50% staining. Statistical analysis: samples with 0/+ staining were considered in low VEGF expression, while samples with ++/+++ expression were considered as high VEGF expression. For DII40 well demarcated vessels showing positivity in endothelial cells were counted per 10hpf after selecting hot spots at scanner view. For statistical analysis, 0 was considered as no expression and 1/10HPF was considered as high expression. Association between levels of VEGF and D II 40 was examined by the two‐way scatter plot and a linear regression analysis of DII 40 over VEGF. The association of positivity between the two markers was assessed using Chi Square test.ResultsHigh VEGF expression was seen 33 of the 43 evaluable patients (76.74%%; 95% CI 61.37%‐88.24%) and high DII 40 expression was seen in 30 of the 44 patients (68.18%; 95% CI 52.42%‐81.39%). Of the 33 patients with high VEGF expression, DII 40 was expressed in 26 of them (78.79%). There was a strong association between VEGF expression and DII 40 expression (p&lt;0.01) (Figure 1). 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 the expression of VEGF and D II 40 was compared with tumor characteristics, there was statistically significant correlation with lymphovascular invasion (LVI) but not with tumor size, type, grade, ER/PR status, Her2neu status or lymph node involvement.ConclusionVEGF expression is high in most tumors. There is significant association between LVI &amp; lymph vessels detection as studied by D II 40 expression and VEGF expression. Further studies in clinical settings are essential before incorporating VEGF expression for prognostication and customized patient care.

A Nomogram Based on Molecular Biomarkers and Radiomics to Predict Lymph Node Metastasis in Breast Cancer.

BackgroundThe aim of this study was to explore the feasibility and efficacy of a non-invasive quantitative imaging evaluation model to assess the lymphatic metastasis of breast cancer based on a radiomics signature constructed using conventional T1-weighted image (T1WI) enhanced MRI and molecular biomarkers.MethodsPatients with breast cancer diagnosed via lymph biopsies between June 2015 and June 2019 were selected for the study. All patients underwent T1WI contrast-enhancement before treatment; lymph biopsy after surgery; and simultaneous Ki-67, COX-2, PR, Her2 and proliferating cell nuclear antigen detection. All images were imported into ITK-SNAP for whole tumor delineation, and AK software was used for radiomics feature extraction. Next, the radiomics signature Rad-score was constructed after reduction of specific radiomic features. A multiple regression logistic model was built by combining the Rad-score and molecular biomarkers based on the minimum AIC.ResultsIn all, 100 patients were enrolled in this study, including 45 with non-lymph node (LN) metastasis and 55 with LN metastasis. A total of 1,051 texture feature parameters were extracted, and LASSO was used to reduce the dimensionality of the radiomics features. The log(λ) was set to 0.002786, and 19 parameters were retained for the construction of the radiomics tag Rad-score. ROC was used to evaluate the diagnostic efficiency of Rad-score: the area under the ROC curve (AUC) of the Rad-score for identifying non-lymphatic and lymphatic metastases was 0.891 in the training cohort and 0.744 in the validation cohort. With the incorporation of tumor molecular markers, the AUCs of the training cohort and validation cohort of the nomogram were 0.936 and 0.793, respectively, which were notably higher than the AUCs of the clinical parameters in the training and validation cohorts (0.719 and 0.588, respectively).ConclusionThe combined model constructed using the Rad-score and molecular biomarkers can be used as an effective non-invasive method to assess LN metastasis of breast cancer. Furthermore, it can be used to quantitatively evaluate the risk of breast cancer LN metastasis before surgery.

Nectin-4 promotes lymphangiogenesis and lymphatic metastasis in breast cancer by regulating CXCR4-LYVE-1 axis

Tumor-induced lymphangiogenesis promotes tumor progression by generating new lymphatic vessels that helps in tumor dissemination to regional lymph nodes and distant sites. Recently, the role of Nectin-4 in cancer metastasis and angiogenesis has been studied, but its role in lymphangiogenesis is unknown. Here, we systematically delineated the role of Nectin-4 in lymphangiogenesis and its regulation in invasive duct carcinoma (IDC). Nectin-4 expression positively correlated with occurrence risk factors associated with breast cancer (alcohol, smoke, lifestyle habit, etc), CXCR4 expression, and LYVE-1-lymphatic vessel density (LVD). LVD was significantly higher in axillary lymph node (ALN) than primary tumor. Depleting Nectin-4, VEGF-C or both attenuated the important lymphangiogenic marker LYVE-1 expression, tube formation, and migration of ALN derived primary cells. Nectin-4 stimulated the expressions of CXCR4 and CXCL12 under hypoxic conditions in ALN derived primary cells. Further, Nectin-4 augmented expressions of lymphatic metastatic markers (e.g. eNOS, TGF-β, CD-105) and MMPs. Induced expressions of Nectin-4 along with other representative metastatic markers were noted in lymph and blood circulating tumor cells (LCTCs and BCTCs) of local and distant metastatic samples. Thus, Nectin-4 displayed a predominant role in promoting tumor-induced lymphangiogenesis and lymphatic metastasis by modulating CXCR4/CXCL12-LYVE-1- axis.

Integrative analyses of scRNA-seq and scATAC-seq reveal CXCL14 as a key regulator of lymph node metastasis in breast cancer.

The potentially different genetics and epigenetics in the primary tumors and metastases affect the efficacy of treatment in breast cancer patients. Nevertheless, the cellular and molecular mechanisms of breast cancer lymph node metastasis still remain elusive. Here, we employed single-cell RNA sequencing to acquire the transcriptomic profiles of individual cells from primary tumors, negative lymph nodes (NLs) and positive lymph nodes (PLs). We also performed a single-cell assay for transposase-accessible chromatin (ATAC) sequencing (scATAC-seq) of the positive and NL samples to get the chromatin accessibility profile. We identified a novel cell subpopulation with an abnormally high expression level of CXCL14 in the PL of breast cancer patients. Cell trajectory analysis also revealed that CXCL14 was increased expressed in the late pseudo-time. Moreover, based on a tissue microarray of 55 patients and the Oncomine database, we validated that CXCL14 expression was significantly higher in breast cancer patients with lymph node metastasis. Furthermore, scATAC-seq identified several transcription factors that may be potential regulation factors for the lymph node metastasis of breast cancer. Thus, our findings will improve our current understanding of the mechanism for lymph node metastasis, and they are potentially valuable in providing novel prognosis markers for the lymphatic metastasis of breast cancer.

Expressions of ZNF436, β-catenin, EGFR, and CMTM5 in breast cancer and their clinical significances.

As the leading malignancy among women, breast cancer is a serious threat to the life and health of women. In this context, it is of particular importance that a proper therapeutic target be identified for breast cancer treatment. We collected the pathological tissues of 80 patients, with the view to discovering appropriate molecular targets for the treatment of breast cancer, this paper analyzes the expressions of ZNF436, β-catenin, EGFR and CMTM5 in breast cancer tissues, as well as their correlations with breast cancer in combination with the clinicopathologic characteristics of studied patients. Immunohistochemistry was utilized to detect the expression levels of ZNF436, β-catenin, EGFR and CMTM5 in cancerous and paracancerous tissues of breast cancer patients. The expression levels of ZNF436, β-Catenin and EGFR in breast cancer tissues were significantly greater than those in paracancerous tissues in this study (p<0.05), while CMTM5 was highly expressed in paracancerous tissues (p<0.05). Additionally, the correlation of the expressions of such indicators with the staging, differentiation and lymphatic metastasis of breast cancer, were also found to be statistically significant at the level p<0.05. The different expression levels of ZNF436, β-catenin, EGFR and CMTM5 in breast cancer and paracancerous tissues open up the possibility of utilizing them as molecular markers for breast cancer. These findings provide a theoretical basis for targeted molecular therapies for breast cancer, and hence carry a significant practical significance.

Studying Lymphatic Metastasis in Breast Cancer: Current Models, Strategies, and Clinical Perspectives

Breast cancer is the most commonly diagnosed cancer in women and the second most common cause of cancer-related deaths in the United States. Although early detection has significantly decreased breast cancer mortality, patients diagnosed with distant metastasis still have a very poor prognosis. The most common site that breast cancer spreads to are local lymph nodes. Therefore, the presence of lymph node metastasis remains one of most important prognostic factors in breast cancer patients. Given its significant clinical implications, increased efforts have been dedicated to better understand the molecular mechanism governing lymph node metastasis in breast cancer. The identification of lymphatic-specific biomarkers, including podoplanin and LYVE-1, has propelled the field of lymphatic metastasis forward. In addition, several animal models such as cell line-derived xenografts, patient-derived xenografts, and spontaneous tumor models have been developed to recreate the process of lymphatic metastasis. Moreover, the incorporation of various -omic platforms have provided further insight into the genetic drivers facilitating lymphatic metastasis, as well as potential biomarkers and therapeutic targets. Here, we highlight various models of lymphatic metastasis, their potential pitfalls, and other tools available to study lymphatic metastasis including imaging modalities and -omic studies.

Macphatics and PoEMs in Postpartum Mammary Development and Tumor Progression.

Postpartum mammary gland involution is a mammalian tissue remodeling event that occurs after pregnancy and lactation to return the gland to the pre-pregnant state. This event is characterized by apoptosis and lysosomal-mediated cell death of the majority of the lactational mammary epithelium, followed by remodeling of the extracellular matrix, influx of immune cell populations (in particular, T helper cells, monocytes, and macrophages), and neo-lymphangiogenesis. This postpartum environment has been shown to be promotional for tumor growth and metastases and may partially account for why women diagnosed with breast cancer during the postpartum period or within 5years of last childbirth have an increased risk of developing metastases when compared to their nulliparous counterparts. The lymphatics and macrophages present during mammary gland involution have been implicated in promoting the observed growth and metastasis. Of importance are the macrophages, which are of the "M2" phenotype and are known to create a pro-tumor microenvironment. In this report, we describe a subset of postpartum macrophages that express lymphatic proteins (PoEMs) and directly interact with lymphatic vessels to form chimeric vessels or "macphatics". Additionally, these PoEMs are very similar to tumor-associated macrophages that also express lymphatic proteins and are present at the sites of lymphatic vessels where tumors escape the tissue and enter the lymphatic vasculature. Further characterizing these PoEMs may offer insight in preventing lymphatic metastasis of breast cancer, as well as provide information for how developmental programming of lymphatic endothelial cells and macrophages can contribute to different disease progression.

Prognostic value of biomarkers EpCAM and \u03b1B-crystallin associated with lymphatic metastasis in breast cancer by iTRAQ analysis

BackgroundMetastasis is responsible for the majority of deaths in a variety of cancer types, including breast cancer. Although several factors or biomarkers have been identified to predict the outcome of patients with breast cancer, few studies have been conducted to identify metastasis-associated biomarkers.MethodsQuantitative iTRAQ proteomics analysis was used to detect differentially expressed proteins between lymph node metastases and their paired primary tumor tissues from 23 patients with metastatic breast cancer. Immunohistochemistry was performed to validate the expression of two upregulated (EpCAM, FADD) and two downregulated (NDRG1, αB-crystallin) proteins in 190 paraffin-embedded tissue samples. These four proteins were further analyzed for their correlation with clinicopathological features in 190 breast cancer patients.ResultsWe identified 637 differentially regulated proteins (397 upregulated and 240 downregulated) in lymph node metastases compared with their paired primary tumor tissues. Data are available via ProteomeXchange with identifier PXD013931. Furthermore, bioinformatics analysis using GEO profiling confirmed the difference in the expression of EpCAM between metastases and primary tumors tissues. Two upregulated (EpCAM, FADD) and two downregulated (NDRG1, αB-crystallin) proteins were associated with the progression of breast cancer. Obviously, EpCAM plays a role in the metastasis of breast cancer cells to the lymph node. We further identified αB-crystallin as an independent biomarker to predict lymph node metastasis and the outcome of breast cancer patients.ConclusionWe have identified that EpCAM plays a role in the metastasis of breast cancer cells to the lymph node. αB-crystallin, a stress-related protein that has recently been shown to be important for cell invasion and survival, was identified as a potential prognostic biomarker to predict the outcome of breast cancer patients.

PTEN and SHIP: Impact on lymphatic metastasis in breast cancer.

Lymph node metastasis is the most common form of metastasis in breast cancer and a crucial indicator influencing breast cancer treatment results. The biological process of lymph node metastasis involves deficiency and mutation of tumor suppressor genes. PTEN and SHIP are critical indicators used to detect occurrence, development, invasion, and metastasis of breast cancer. Loss of expressions of PTEN and SHIP may contribute to lymphatic metastasis of breast cancer, so they can be used as objective indicators for judging the biological behavior of breast cancer. In this study, we perform a comprehensive analysis to investigate the effect of PTEN and SHIP gene expression on regulating lymph node metastasis in breast cancer.

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

PGE2 promotes breast cancer-associated lymphangiogenesis by activation of EP4 receptor on lymphatic endothelial cells

BackgroundLymphatic metastasis, facilitated by lymphangiogenesis is a common occurrence in breast cancer, the molecular mechanisms remaining incompletely understood. We had earlier shown that cyclooxygenase (COX)-2 expression by human or murine breast cancer cells promoted lymphangiogenesis and lymphatic metastasis by upregulating VEGF-C/D production by tumor cells or tumor-associated macrophages primarily due to activation of the prostaglandin receptor EP4 by endogenous PGE2. It is not clear whether tumor or host-derived PGE2 has any direct effect on lymphangiogenesis, and if so, whether EP4 receptors on lymphatic endothelial cells (LEC) play any role.MethodsHere, we address these questions employing in vitro studies with a COX-2-expressing and VEGF-C/D-producing murine breast cancer cell line C3L5 and a rat mesenteric (RM) LEC line and in vivo studies in nude mice.ResultsRMLEC responded to PGE2, an EP4 agonist PGE1OH, or C3L5 cell-conditioned media (C3L5-CM) by increased proliferation, migration and accelerated tube formation on growth factor reduced Matrigel. Native tube formation by RMLEC on Matrigel was abrogated in the presence of a selective COX-2 inhibitor or an EP4 antagonist. Addition of PGE2 or EP4 agonist, or C3L5-CM individually in the presence of COX-2 inhibitor, or EP4 antagonist, restored tube formation, reinforcing the role of EP4 on RMLEC in tubulogenesis. These results were partially duplicated with a human dermal LEC (HMVEC-dLyAd) and a COX-2 expressing human breast cancer cell line MDA-MB-231. Knocking down EP4 with shRNA in RMLEC abrogated their tube forming capacity on Matrigel in the absence or presence of PGE2, EP4 agonist, or C3L5-CM. RMLEC tubulogenesis following EP4 activation by agonist treatment was dependent on PI3K/Akt and Erk signaling pathways and VEGFR-3 stimulation. Finally in a directed in vivo lymphangiogenesis assay (DIVLA) we demonstrated the lymphangiogenic as well as angiogenic capacity of PGE2 and EP4 agonist in vivo.Discussion/conclusionsThese results demonstrate the roles of tumor as well as host-derived PGE2 in inducing lymphangiogenesis, at least in part, by activating EP4 and VEGFR-3 on LEC. EP4 being a common target on both tumor and host cells contributing to tumor-associated lymphangiogenesis reaffirms the therapeutic value of EP4 antagonists in the intervention of lymphatic metastasis in breast cancer.

Relationship between peritumoral lymphatic microvessel density and the clinical and pathological characteristics of invasive breast cancer

Abstract Objective The aim of the study was to determine the morphological characteristics of lymphatic microvessels and the relationship between lymphatic microvessel density (LMVD) and clinical and pathological characteristics of invasive breast cancer. Methods Tissue specimens and clinical pathological data of 51 cases of female breast cancer were collected in the General Hospital of Shenyang Military Region (Shenyang, China) from January 2007 to October 2011. Another 20 breast fibroadenoma tissue samples were used as controls. All specimens were cut into 4-μm slices, and immunohistochemically stained using streptomycin-resistant avidin peroxidase antibody D2-40. SPSS 17.0 for Windows was used to perform all analyses. Results A total of 38 breast cancer tissue specimens showed varied staining with monoclonal antibody D2-40. The rate of positive staining was in these tissues was 74.5% (38/51), which is significantly higher than that observed in breast fibroadenoma tissues (chi-square = 35.197, P = 0.000). The average LMVD in 38 cases of breast cancer was (26.46 ± 10.06) microvessels/100× magnification field, which was higher than that in the control group (t = 10.74, P = 0.000). Microvessels in peritumoral tissues were abundant, with an average LMVD of (38.42 ± 11.38) microvessels/100× magnification field. Based on layered analysis, the expression level of peritumoral LMVD was correlated with metastasis of lymph nodes, tumor size, and the expression of C-erbB-2 (P &lt; 0.05); however, there was no correlation with age or expression of estrogen receptors or progesterone receptors (P &gt; 0.05). Conclusion Lymphatic microvessels detected using D2-40 antibody are mainly present in the peritumoral region of breast cancer tissues, and LMVD showed a correlation with lymph node metastasis and the expression of C-erbB-2. Positive lymphatic vessels, especially in the peritumoral region, may provide a path for lymphatic metastasis in breast cancer. Peritumoral LMVD may be used to estimate the prognosis of patients with breast cancer and may aid in research on treatment methods.

Breast cancer sphingosine-1-phosphate is associated with phospho-sphingosine kinase 1 and lymphatic metastasis

BackgroundSphingosine-1-phosphate (S1P), a pleiotropic bioactive lipid mediator, has been implicated as a key regulatory molecule in cancer through its ability to promote cell proliferation, migration, angiogenesis, and lymphangiogenesis. Previous studies suggested that S1P produced by sphingosine kinase 1 (SphK1) in breast cancer plays important roles in progression of disease and metastasis. However, the associations between S1P and clinical parameters in human breast cancer have not been well investigated to date. Materials and methodsWe determined levels of S1P and other sphingolipids in breast cancer tissue by electrospray ionization-tandem mass spectrometry. Associations between S1P levels and clinicopathologic features of the tumors were analyzed. Expression of phospho-SphK1 (pSphK1) in breast cancer tissues was determined by immunohistochemical scoring. ResultsLevels of S1P in breast cancer tissues were significantly higher in patients with high white blood cell count in the blood than those patients without. S1P levels were lower in patients with human epidermal growth factor receptor 2 overexpression and/or amplification than those patients without. Furthermore, cancer tissues with high pSphK1 expression showed significantly higher levels of S1P than cancer tissues without. Finally, patients with lymph node metastasis showed significantly higher levels of S1P in tumor tissues than the patients with negative nodes. ConclusionsTo our knowledge, this is the first study to demonstrate that high expression of pSphK1 is associated with higher levels of S1P, which in turn is associated with lymphatic metastasis in breast cancer.

Ki-67 expression in luminal type breast cancer and its association with the clinicopathology of the cancer.

The aim of the present study was to examine Ki-67 expression in tissue of luminal type breast cancer and to investigate the association between the Ki-67 expression level and clinicopathology of breast cancer. A total of 62 patients with breast cancer were included in the study group, whereas 30 patients undergoing a health check-up who were diagnosed with breast hyperplasia were included in the control group. Levels of Ki-67 expression in patients of the two groups were assessed using fluorescent quantitative polymerase chain reaction. The association between Ki-67 and the clinicopathology of the cancer was investigated by analyzing cancer cell proliferation and migration by reducing Ki-67 expression in the human MCF-7 cancer cell line. Compared with the controls, Ki-67 expression was significantly increased in the serum and cancer tissue of breast cancer patients (P<0.05). Ki-67 mRNA expression was significantly higher in cancer tissue than that in the corresponding paracancerous tissue of breast cancer (P<0.05). In addition, a high expression of Ki-67 was positively correlated with the clinical staging of tumor, tumor size and lymphatic metastasis of breast cancer, with statistical significance (P<0.05). In MCF-7 cells with a reduced Ki-67 expression, the proliferation activity and migration of breast cancer cells were significantly reduced (P<0.05). In conclusion, Ki-67 may be involved in promoting the genesis and development of breast cancer by affecting the proliferation and migration of cancer cells.

Proteomic analysis of O-GlcNAcylated proteins in invasive ductal breast carcinomas with and without lymph node metastasis

The potential role of protein O-GlcNAcylation in cancer has been studied extensively, and the spread of cancer cells to regional lymph nodes is the first step in the dissemination of breast cancer. However, the correlation between O-GlcNAcylation and lymphatic metastasis in breast cancer remains elusive. In this study, we demonstrated that the overall O-GlcNAcylation as well as O-GlcNAc transferase (OGT) tends to decrease in response to the augmentation of lymph node metastasis (LNM) in invasive ductal breast carcinomas (IDCs). Although accumulating evidence indicates that individual O-GlcNAcylation may be important in the pathogenesis of breast cancer, O-GlcNAcylated proteins in IDCs are still largely unexplored. Herein, O-GlcNAcylated proteins of IDCs were chemo-enzymatically enriched and identified via liquid chromatography combined with tandem mass spectrometry. In total, 155 O-GlcNAcylated proteins were determined, of which 41 were only observed in LNM tissues, while 40 were unique in non-LNM samples. Gene ontology analysis showed that O-GlcNAc is primarily a nucleocytoplasmic post-translational modification, and most enriched functional terms were related to cancer development in both metastatic and non-metastatic IDCs. Moreover, several O-GlcNAcylated proteins involved in glycolysis and its accessory pathway were identified from LNM and non-LNM groups, respectively. These results indicate that the O-GlcNAcylation statuses of individual proteins were independent of the overall O-GlcNAcylation levels of metastatic and non-metastatic IDCs. Aberrant O-GlcNAc modification of these proteins might be associated with LNM progression.

HtrA3 is negatively correlated with lymph node metastasis in invasive ductal breast cancer

Breast cancer is the most common cancer in women worldwide. Studies have shown that the high temperature requirement factor A3 (HtrA3) is involved in important physiological processes including maintenance of mitochondrial homeostasis, cell death, and cell signaling. HtrA3 is reported to be downregulated in several cancers and has been correlated with advancing cancer stage. We performed a retrospective study using our breast cancer tissue bank to investigate whether the expression of HtrA3 correlated with lymphatic metastasis in breast cancer and whether the expression of HtrA3 was correlated with estrogen receptor (ER) and progesterone receptor (PR) expression in breast cancer. Breast cancer tissues from 156 invasive ductal breast cancer patients with or without lymphatic metastasis were collected and the levels of HtrA3 were measured by immunohistochemistry and western blotting. The expression of HtrA3 was lower in breast cancer. In particular, HtrA3 expression in breast cancer with lymphatic metastasis was lower than that in breast cancer without lymphatic metastasis. In breast cancers with no lymphatic metastasis, the expression of HtrA3 was lower in patients with ER- and PR-positive tumors, but when breast cancers with lymphatic metastasis were analyzed, there was no difference in HtrA3 expression between ER- and PR-positive or ER- and PR-negative tumors. These data suggest that the expression of HtrA3 was negatively correlated with lymphatic metastasis in breast cancer but not correlated with ER and PR positivity or negativity. A better understanding of the mechanism of HtrA3 may provide the basis for future development of a novel therapeutic target in breast cancer.

A potential association between the number of CA repeats in the promoter region of the ADAMTS9 gene with lymphatic metastasis of breast cancer

We investigated the effect of the number of cytosine-adenine (CA) repeats in the ADAMTS9 promoter region on breast cancer lymphatic metastasis. Materials and methods: Thirty-one postoperative breast cancer patients were selected and examined retrospectively. The patients were classified into 2 groups: metastatic or nonmetastatic. Thirty healthy women were selected as the control group, and their peripheral blood was obtained. Following DNA isolation from the cancer tissue specimens and peripheral blood, the promoter region of the ADAMTS9 gene was directly sequenced and the number of CA repeats was determined. Results: The number of CA repeats ranged between 19 and 21 in the control and metastatic groups. However, in the nonmetastatic group, the number of CA repeats ranged between 17 and 18. This difference in the median number of CA repeats between the control group and the nonmetastatic group was statistically significant. Conclusion: A potential relationship may exist between lymphatic metastasis in breast cancer and the number of CA repeats in the promoter region of the ADAMTS9 gene. Our study indicates a potential association between the number of CA microsatellite repeats in the promoter region of the ADAMTS9 gene and breast cancer lymphatic metastasis.