LncRNA SEMA3B-AS1 suppresses the tumor-initiating characteristics of triple negative breast cancer via engaging in MLL4-mediated H3K4 trimethylation.

Long noncoding RNAs (lncRNAs) are crucial regulators of tumor-initiating cells (TICs) and hold particular importance in triple negative breast cancer (TNBC). Yet, the precise mechanisms by which TIC-associated lncRNAs influence TNBC remain unclear. Our research utilized The Cancer Genome Atlas Breast Cancer (BC) data set to identify prognostic lncRNAs. We then conducted extensive assays to explore their impact on the tumor-initiating phenotype of TNBC cells and the underlying mechanisms. Notably, we found that low expression of lncRNA SEMA3B-AS1 correlated with unfavorable survival in BC patients. SEMA3B-AS1 was also downregulated in TNBC and linked to advanced tumor stage. Functional experiments confirmed its role as a TIC-suppressing lncRNA, curtailing mammosphere formation, ALDH + TIC cell proportion, and impairing clonogenicity, migration, and invasion. Mechanistic insights unveiled SEMA3B-AS1's nuclear localization and interaction with MLL4 (mixed-lineage leukemia 4), triggering H3K4 methylation-associated transcript activation and thus elevating the expression of SEMA3B, a recognized tumor suppressor gene. Our findings emphasize SEMA3B-AS1's significance as a TNBC-suppressing lncRNA that modulates TIC behavior. This study advances our comprehension of lncRNA's role in TNBC progression, advocating for their potential as therapeutic targets in this aggressive BC subtype.

Effect, sensitivity, specificity and accuracy of ultrasonic assessment of axillary lymph node-negative breast cancer.

Objective: To investigate the diagnostic value of ultrasound for patients with axillary lymph node-negative breast cancer (ALNNBC). Methods: A retrospective analysis was performed on the clinical data of 204 breast cancer patients who were admitted by Quanzhou First Hospital Affiliated to Fujian Medical University between October 2020 and May 2022. According to the results of axillary lymph node (ALN) examination, the patients were assigned to a positive group(n=102) and a negative group(n=102). All patients underwent diagnosis with color Doppler ultrasound, with pathological diagnosis as the "gold standard" to determine the sensitivity and specificity of ultrasonic diagnosis. A receiver operating characteristic(ROC) curve was established to analyze the efficiency of ultrasonic diagnosis and compare the ultrasonographic features and flow grades between the two groups. Results: Differences were statistically significant between the two groups in ultrasonographic features of lesions(negative vs positive, all p<0.05), including morphological irregularity(59.8% vs 85.3%), spiky margins(19.6% vs 63.7%), posterior echo attenuation(19.6% vs 44.1%) and microcalcification(40.2% vs 55.89%). The negative group had a lower proportion of patients with grade 2-3 ultrasound blood flow when compared with the positive group(32.4% vs 56.86%), and the difference was statistically significant(p<0.05). Ultrasonic diagnosis of ALNNBC had a sensitivity of 88.24%(90/102), a specificity of 92.16%(94/102), a coincidence rate of 90.20% (184/204), a 95% CI of 0.845-0.928, and an AUC of 0.879. Conclusions: Ultrasonic diagnosis of ALNNBC is relatively efficient as ultrasonographic features and ultrasound blood flow signals can provide a scientific basis for the diagnosis of ALNNBC.

ZNF703 promotes triple-negative breast cancer cells through cell-cycle signaling and associated with poor prognosis.

BACKGROUND: The oncogenic drivers of triple-negative breast cancer (TNBC), which is characterized by worst prognosis compared with other subtypes, are poorly understood. Although next-generation sequencing technology has facilitated identifying potential targets, few of the findings have been translated into daily clinical practice. The present study is aimed to explore ZNF703 (Zinc finger 703) function and its underlying mechanism in TNBC. METHODS: ZNF703 expressions in tissue microarray were retrospectively examined by immunohistochemistry. The cell proliferation by SRB assay and colony formation assay, as well as cell cycle distribution by flow cytometry were assessed. The protein levels associated with possible underlying molecular mechanisms were evaluated by western blotting. Kaplan-Meier analysis was used to plot survival analysis. RESULTS: Our data suggest that ZNF703 expressed in 34.2% of triple-negative human breast tumors by immunohistochemistry. In vitro, ZNF703 knockdown had potent inhibitory effects on TNBC cell proliferation and cell cycle, with cyclin D1, CDK4, CDK6, and E2F1 downregulated, while Rb1 upregulated. Moreover, Kaplan-Meier analysis showed that high mRNA expression of ZNF703 was correlated to worse overall survival (HR for high expression was 3.04; 95% CI, 1.22 to 7.57, P = 0.017). CONCLUSIONS: Taken together, the results identified that targeting ZNF703 contributed to the anti-proliferative effects in TNBC cells, due to induced G1-phase arrest. This study is the first to identify ZNF703 as a potentially important protein that is involved in TNBC progression.

Validation of the Eighth American Joint Committee on Cancer Anatomic and Prognostic Staging System for Breast Cancer.

BACKGROUND: The eighth edition of new staging systems for breast cancer incorporated four biological factors and the anatomic staging system. Validating analysis on Chinese patients has been limited. Our study performed analysis comparing the prognostic value of the staging system based on Chinese data. METHODS AND MATERIALS: All patients were classified according to the eighth edition and compared between anatomic and prognostic staging systems. The Kaplan-Meier test was used to calculate the overall survival (OS) and disease-free survival (DFS). We performed Harrell concordance index (C-index) analyses to quantify a models' predictive performance. Akaike information criterion (AIC) via Cox regression analysis was used to conduct bootstrap-based goodness-of-fit comparisons of the competing staging systems. RESULTS: A total of 1556 patients were enrolled in the cohort. The median follow-up time was 76 mo (range, 4-146 mo), the median age was 48 y old (range, 21-87 y). The ratio of movement between anatomic stage (AS) and prognostic stage (PS) was 50.9%. Of these, 691 (44.5%) AS patients were down staged and 100 (6.4%) patients were upstaged when reclassified based on PS. Significant differences between two stages were achieved for stage IIIC in 5-y OS rates and for IIIB in 5-y DFS rates (63.5% versus 50.0% and 58.0% versus44.0%). The value of the C-index for PS and AS were 0.711 and 0.687 (P = 0.04). The AIC reaches a value of 3452.9 for the PS and a value of 3476.4 for the AS. CONCLUSIONS: The PS might provide better accuracy than the AS in predicting the prognosis of Chinese female breast cancer patients. It also provides a strong basis for the utility of clinical biomarkers to evaluate the prognosis of patients.

Single molecule chromogenic in situ hybridization assay for RNA visualization in fixed cells and tissues.

Visualization of gene expression at single RNA molecular level represents a great challenge to both imaging technologies and molecular engineering. Here we show a single molecule chromogenic in situ hybridization (smCISH) assay that enables counting and localizing individual RNA molecules in fixed cells and tissue under bright-field microscopy. Our method is based on in situ padlock probe assays directly using RNA as a ligation template and rolling circle amplification combined with enzyme catalyzed chromogenic reaction for amplification product visualization. We show potential applications of our method by detecting gene expression variations in single cells, subcellular localization information of expressed genes, and gene expression heterogeneity in formalin-fixed, paraffin-embedded tissue sections. This facile and straightforward method can in principle be applied to any type of RNA molecules in different samples.

Comparative analysis of surgical modalities for the gynecomastia treatment: efficiency and aesthetic outcomes.

This retrospective study rigorously compares the clinical efficacy of three surgical methodologies for treating gynecomastia while providing guidance for future surgical modality selection. We analyzed records of 77 gynecomastia patients treated between January 2015 and October 2022. Patients were categorized into three groups: Group A (subcutaneous gland resection via areola incision), Group B (liposuction combined with single-hole endoscopic gland resection), and Group C (liposuction combined with three-hole endoscopic gland resection). Parameters assessed included patient demographics, intraoperative bleeding, surgical duration, hospitalization duration, costs, postoperative drainage, complications, and patient satisfaction. Group A had significantly shorter operation time and lower cost than Groups B and C (P < 0.05). There were no significant differences in postoperative drainage (P > 0.05). Group A had a higher incidence of subcutaneous fluid complications. All groups achieved 100% overall postoperative efficiency. Group B demonstrated superior outcomes for scarring and patient satisfaction. All three surgical modalities effectively treat gynecomastia. Circumareolar incision subcutaneous gland resection is optimal for mild to moderate cases due to reduced operation time and cost. Liposuction with single-hole endoscopic gland resection and three-hole endoscopic gland resection offers fewer complications and discreet incisions. Notably, the liposuction and single-hole endoscopic approach yielded superior postoperative patient satisfaction, aligning with minimally invasive principles and warranting broad clinical application.

A ceRNA network-mediated over-expression of cuproptosis-related gene SLC31A1 correlates with poor prognosis and positive immune infiltration in breast cancer.

Introduction: Solute carrier family 31 member 1(SLC31A1) has been reported as the copper importer, and was identified to be involved in the process of "cuproptosis". However, the mechanism of SLC31A1 in breast cancer remains unclear. Methods: We examined the expression of SLC31A1 mRNA in breast cancer tissues and cell lines using Real-time PCR. The data for this study were obtained from The Cancer Genome Atlas (TCGA) database and analyzed via R 3.6.3. TIMER, UALCAN, GEPIA2, STRING, Metascape, Kaplan-Meier Plotter, starBase and miRNet websites were used for a comprehensive analysis of SLC31A1. Results: Our study suggested that SLC31A1 mRNA was over-expressed in breast tumor tissue and breast cancer cell lines, and which was closely related to poor relapse-free survival (RFS) and distant metastasis-free survival (DMFS). In addition, we constructed a co-expression network of SLC31A1. Functional enrichment analysis indicated that they were mainly involved in copper ion transport. Interestingly, SLC31A1 expression was positively associated with all m6A-related genes, especially with YTHDF3 (r = 0.479). Importantly, the LINC00511/miR-29c-3p/SLC31A1 axis was identified as the most potential pathway promoting breast cancer progress by affecting copper transport. Furthermore, the expression level of SLC31A1 in breast cancer was positively correlated with tumor immune cell infiltration, immune cell biomarkers and cancer-associated fibroblast (CAF). Conclusion: Up-regulation of SLC31A1 expression and regulation of copper ion transport mediated by LINC00511-miR-29-3p axis is related to poor prognosis and positively correlated with tumor immune infiltration in breast cancer.

Combined signature of N7-methylguanosine regulators with their related genes and the tumor microenvironment: a prognostic and therapeutic biomarker for breast cancer.

Background: Identifying predictive markers for breast cancer (BC) prognosis and immunotherapeutic responses remains challenging. Recent findings indicate that N7-methylguanosine (m7G) modification and the tumor microenvironment (TME) are critical for BC tumorigenesis and metastasis, suggesting that integrating m7G modifications and TME cell characteristics could improve the predictive accuracy for prognosis and immunotherapeutic responses. Methods: We utilized bulk RNA-sequencing data from The Cancer Genome Atlas Breast Cancer Cohort and the GSE42568 and GSE146558 datasets to identify BC-specific m7G-modification regulators and associated genes. We used multiple m7G databases and RNA interference to validate the relationships between BC-specific m7G-modification regulators (METTL1 and WDR4) and related genes. Single-cell RNA-sequencing data from GSE176078 confirmed the association between m7G modifications and TME cells. We constructed an m7G-TME classifier, validated the results using an independent BC cohort (GSE20685; n = 327), investigated the clinical significance of BC-specific m7G-modifying regulators by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis, and performed tissue-microarray assays on 192 BC samples. Results: Immunohistochemistry and RT-qPCR results indicated that METTL1 and WDR4 overexpression in BC correlated with poor patient prognosis. Moreover, single-cell analysis revealed relationships between m7G modification and TME cells, indicating their potential as indicators of BC prognosis and treatment responses. The m7G-TME classifier enabled patient subgrouping and revealed significantly better survival and treatment responses in the m7Glow+TMEhigh group. Significant differences in tumor biological functions and immunophenotypes occurred among the different subgroups. Conclusions: The m7G-TME classifier offers a promising tool for predicting prognosis and immunotherapeutic responses in BC, which could support personalized therapeutic strategies.

A novel disulfidptosis-associated expression pattern in breast cancer based on machine learning.

Background: Breast cancer (BC), the leading cause of cancer-related deaths among women, remains a serious threat to human health worldwide. The biological function and prognostic value of disulfidptosis as a novel strategy for BC treatment via induction of cell death remain unknown. Methods: Gene mutations and copy number variations (CNVs) in 10 disulfidptosis genes were evaluated. Differential expression, prognostic, and univariate Cox analyses were then performed for 10 genes, and BC-specific disulfidptosis-related genes (DRGs) were screened. Unsupervised consensus clustering was used to identify different expression clusters. In addition, we screened the differentially expressed genes (DEGs) among different expression clusters and identified hub genes. Moreover, the expression level of DEGs was detected by RT-qPCR in cellular level. Finally, we used the least absolute shrinkage and selection operator (LASSO) regression algorithm to establish a prognostic feature based on DEGs, and verified the accuracy and sensitivity of its prediction through prognostic analysis and subject operating characteristic curve analysis. The correlation of the signature with the tumor immune microenvironment and tumor stemness was analyzed. Results: Disulfidptosis genes showed significant CNVs. Two clusters were identified based on three DRGs (DNUFS1, LRPPRC, SLC7A11). Cluster A was found to be associated with better survival outcomes(p < 0.05) and higher levels of immune cell infiltration(p < 0.05). A prognostic signature of four disulfidptosis-related DEGs (KIF21A, APOD, ALOX15B, ELOVL2) was developed by LASSO regression analysis. The signature showed a good prediction ability. In addition, the prognostic signature in this study were strongly related to the tumor microenvironment (TME), tumor immune cell infiltration, tumor mutation burden (TMB), tumor stemness, and drug sensitivity. Conclusion: The prognostic signature we constructed based on disulfidptosis-DEGs is a good predictor of prognosis in patients with BC. This prognostic signature is closely related to TME, and its potential correlation provides clues for further studies.

Development of a risk-stratification scoring system for predicting risk of breast cancer based on non-alcoholic fatty liver disease, non-alcoholic fatty pancreas disease, and uric acid.

Many breast cancer patients have both non-alcoholic fatty liver disease (NAFLD) and non-alcoholic fatty pancreas disease (NAFPD). Consequently, we hypothesized that NAFPD and NAFLD were associated with breast cancer, and aimed to build a novel risk-stratification scoring system based on it. In this study, a total of 961 patients with breast cancer and 1,006 non-cancer patients were recruited. The clinical characteristics were collected and analyzed using logistic analysis. Risk factors were assessed by a risk rating system. Univariate analysis showed that body mass index, triglyceride, total cholesterol, NAFLD, NAFPD, low-density lipoprotein, and uric acid (UA) were significantly related to breast cancer. Among them, NAFLD, NAFPD, and UA were independent risk factors related to breast cancer identified by multivariate analysis. The risk assessment model was established based on these factors and demonstrated that the odds ratio sharply increased with the rising scores. Compared with the low-risk group, the odds ratio in the intermediate- and high-risk groups were 1.662 (1.380-2.001) and 3.185 (2.145-4.728), respectively. In conclusion, the risk-stratification scoring system combining NAFLD, NAFPD, and UA can accurately predict the occurrence of breast cancer.

Construction of Prognostic Risk Model of 5-Methylcytosine-Related Long Non-Coding RNAs and Evaluation of the Characteristics of Tumor-Infiltrating Immune Cells in Breast Cancer.

Purpose: The role of 5-methylcytosine-related long non-coding RNAs (m5C-lncRNAs) in breast cancer (BC) remains unclear. Here, we aimed to investigate the prognostic value, gene expression characteristics, and correlation between m5C-lncRNA risk model and tumor immune cell infiltration in BC. Methods: The expression matrix of m5C-lncRNAs in BC was obtained from The Cancer Genome Atlas database, and the lncRNAs were analyzed using differential expression analysis as well as univariate and multivariate Cox regression analysis to eventually obtain BC-specific m5C-lncRNAs. A risk model was developed based on three lncRNAs using multivariate Cox regression and the prognostic value, accuracy, as well as reliability were verified. Gene set enrichment analysis (GSEA) was used to analyze the Kyoto Encyclopedia of Genes and Genomes signaling pathway enrichment of the risk model. CIBERSORT algorithm and correlation analysis were used to explore the characteristics of the BC tumor-infiltrating immune cells. Finally, reverse transcription-quantitative polymerase chain reaction was performed to detect the expression level of three lncRNA in clinical samples. Results: A total of 334 differential m5C-lncRNAs were identified, and three BC-specific m5C-lncRNAs were selected, namely AP005131.2, AL121832.2, and LINC01152. Based on these three lncRNAs, a highly reliable and specific risk model was constructed, which was proven to be closely related to the prognosis of patients with BC. Therefore, a nomogram based on the risk score was built to assist clinical decisions. GSEA revealed that the risk model was significantly enriched in metabolism-related pathways and was associated with tumor immune cell infiltration based on the analysis with the CIBERSORT algorithm. Conclusion: The efficient risk model based on m5C-lncRNAs associated with cancer metabolism and tumor immune cell infiltration could predict the survival prognosis of patients, and AP005131.2, AL121832.2, and LINC01152 could be novel biomarkers and therapeutic targets for BC.

Comparison of the Gut Microbiota in Patients with Benign and Malignant Breast Tumors: A Pilot Study.

The microbiome plays diverse roles in many diseases and can potentially contribute to cancer development. Breast cancer is the most commonly diagnosed cancer in women worldwide. Thus, we investigated whether the gut microbiota differs between patients with breast carcinoma and those with benign tumors. The DNA of the fecal microbiota community was detected by Illumina sequencing and the taxonomy of 16S rRNA genes. The α-diversity and ß-diversity analyses were used to determine richness and evenness of the gut microbiota. Gene function prediction of the microbiota in patients with benign and malignant carcinoma was performed using PICRUSt. There was no significant difference in the α-diversity between patients with benign and malignant tumors (P = 3.15e-1 for the Chao index and P = 3.1e-1 for the ACE index). The microbiota composition was different between the 2 groups, although no statistical difference was observed in ß-diversity. Of the 31 different genera compared between the 2 groups, level of only Citrobacter was significantly higher in the malignant tumor group than that in benign tumor group. The metabolic pathways of the gut microbiome in the malignant tumor group were significantly different from those in benign tumor group. Furthermore, the study establishes the distinct richness of the gut microbiome in patients with breast cancer with different clinicopathological factors, including ER, PR, Ki-67 level, Her2 status, and tumor grade. These findings suggest that the gut microbiome may be useful for the diagnosis and treatment of malignant breast carcinoma.

Prognostic Significance and Tumor Immune Microenvironment Heterogenicity of m5C RNA Methylation Regulators in Triple-Negative Breast Cancer.

PURPOSE: The m5C RNA methylation regulators are closely related to tumor proliferation, occurrence, and metastasis. This study aimed to investigate the gene expression, clinicopathological characteristics, and prognostic value of m5C regulators in triple-negative breast cancer (TNBC) and their correlation with the tumor immune microenvironment (TIM). METHODS: The TNBC data, Luminal BC data and HER2 positive BC data set were obtained from The Cancer Genome Atlas and Gene Expression Omnibus, and 11 m5C RNA methylation regulators were analyzed. Univariate Cox regression and the least absolute shrinkage and selection operator regression models were used to develop a prognostic risk signature. The UALCAN and cBioportal databases were used to analyze the gene characteristics and gene alteration frequency of prognosis-related m5C RNA methylation regulators. Gene set enrichment analysis was used to analyze cellular pathways enriched by prognostic factors. The Tumor Immune Single Cell Hub (TISCH) and Timer online databases were used to explore the relationship between prognosis-related genes and the TIM. RESULTS: Most of the 11 m5C RNA methylation regulators were differentially expressed in TNBC and normal samples. The prognostic risk signature showed good reliability and an independent prognostic value. Prognosis-related gene mutations were mainly amplified. Concurrently, the NOP2/Sun domain family member 2 (NSUN2) upregulation was closely related to spliceosome, RNA degradation, cell cycle signaling pathways, and RNA polymerase. Meanwhile, NSUN6 downregulation was related to extracellular matrix receptor interaction, metabolism, and cell adhesion. Analysis of the TISCH and Timer databases showed that prognosis-related genes affected the TIM, and the subtypes of immune-infiltrating cells differed between NSUN2 and NSUN6. CONCLUSION: Regulatory factors of m5C RNA methylation can predict the clinical prognostic risk of TNBC patients and affect tumor development and the TIM. Thus, they have the potential to be a novel prognostic marker of TNBC, providing clues for understanding the RNA epigenetic modification of TNBC.

Identification of Seven Cell Cycle-Related Genes with Unfavorable Prognosis and Construction of their TF-miRNA-mRNA regulatory network in Breast Cancer.

Breast cancer (BC), with complex tumorigenesis and progression, remains the most common malignancy in women. We aimed to explore some novel and significant genes with unfavorable prognoses and potential pathways involved in BC initiation and progression via bioinformatics methods. BC tissue-specific microarray datasets of GSE42568, GSE45827 and GSE54002, which included a total of 651 BC tissues and 44 normal breast tissues, were obtained from the Gene Expression Omnibus (GEO) database, and 124 differentially expressed genes (DEGs) were identified between BC tissues and normal breast tissues via R software and an online Venn diagram tool. Database for Annotation, Visualization and Integration Discovery (DAVID) software showed that 65 upregulated DEGs were mainly enriched in the regulation of the cell cycle, and Search Tool for the Retrieval of Interacting Genes (STRING) software identified the 39 closest associated upregulated DEGs in protein-protein interactions (PPIs), which validated the high expression of genes in BC tissues by the Gene Expression Profiling Interactive Analysis (GEPIA) tool. In addition, 36 out of 39 BC patients showed significantly worse outcomes by Kaplan-Meier plotter (KM plotter), and an additional Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed that seven genes (cyclin E2 (CCNE2), cyclin B1 (CCNB1), cyclin B2 (CCNB2), mitotic checkpoint serine/threonine kinase B (BUB1B), dual-specificity protein kinase (TTK), cell division cycle 20 (CDC20), and pituitary tumor transforming gene 1 (PTTG1)) were markedly enriched in the cell cycle pathway. Analysis of the clinicopathological characteristics of hub genes revealed that seven cell cycle-related genes (CCRGs) were significantly highly expressed in four BC subtypes (luminal A, luminal B, HER2-positive and triple-negative (TNBC)), and except for the CCNE2 gene, high expression levels were significantly associated with tumor pathological grade and stage and metastatic events of BC. Furthermore, genetic mutation analysis indicated that genetic alterations of CCRGs could also significantly affect BC patients' prognosis. A quantitative real-time polymerase chain reaction (qRT-PCR) assay found that the seven CCRGs were significantly differentially expressed in BC cell lines. Integration of published multilevel expression data and a bioinformatics computational approach were used to predict and construct a regulation mechanism: a transcription factor (TF)-microRNA (miRNA)-messenger RNA (mRNA) regulation network. The present work is the first to construct a regulatory network of TF-miRNA-mRNA in BC for CCRGs and provides new insights into the molecular mechanism of BC.

Effect of node status on breast cancer survival by subtype: a single-center retrospective cohort study.

BACKGROUND: Nodal involvement and molecular subtypes were used as independent prognostic indicators in women with breast cancer. However, they did not adequately address the effect of node status by subtype in outcomes. METHODS: We performed a retrospective review of data from 2004 to 2011 from the Affiliated Union Hospital of Fujian Medical University with newly diagnosed stage I to III breast cancer to investigate the relationship between node status and 5-year disease-free survival (DFS) and breast cancer-specific survival (BCSS) by molecular subtype. The Cox proportional hazards model was used for multivariate analysis. RESULTS: Median follow-up time was 6.4 years. Luminal HER2 and luminal B were the subtypes with a higher percentage of nodal involvement and high-volume nodal involvement (≥4 positive lymph node) than luminal A. The effect of node status on the prognosis varied with molecular subtype. There was no difference in 5-year DFS and BCSS between stage N1 or N2 and N0 groups in patients with luminal A disease. Nodal involvement in women with the luminal B, luminal HER2, and triple-negative subtypes showed significant difference for 5-year DFS and BCSS compared to the node negative group. CONCLUSIONS: Nodal involvement seems to be associated with worse survival in women with the luminal B, luminal HER2, and triple-negative subtypes, but not with the luminal A subtype.

Different prognosis by subtype in the early mucinous breast cancer: a SEER population-based analysis.

BACKGROUND: Our study aims to investigate the clinicopathological characteristics and survival outcomes of mucinous breast cancer (MBC), and explore the effect of histology type on the breast cancer-specific survival (BCSS) by different subtypes. METHODS: we identified 7,083 patients who were diagnosed with MBC and 248,751 with infiltrating ductal carcinoma (IDC) by using the Surveillance, Epidemiology and End Results (SEER) database. The propensity score matching was used to match baseline characteristics among MBC and IDC, and multivariable cox proportional hazards models were used to analyze the relationship between histology type stratified by subtype and BCSS. RESULTS: MBC patients were associated with a fewer nodal involvement, lower grade, earlier stage, more estrogen receptor (ER) or progesterone receptor (PR) positive, and more favorable prognosis compared to the overall IDC population. After 1:1 matching of MBC with IDC by other factors, we found that MBC patients presented better prognosis than the Matched IDC for BCSS. Analysis among ER+PR+ subgroup revealed that MBC patients was significantly better than that Matched IDC patients for BCSS (HR =0.78, 95% CI, 0.63-0.96). However, the survival analysis in the ER+PR- or ER-PR- subgroups suggested that no significant difference was seen between MBC patients and matched IDC patients for BCSS. CONCLUSIONS: Our findings support that MBC seems to be an independent factor for the better prognosis for breast cancer patients with ER+PR+ breast cancer but not in those with ER+PR- or ER-PR- disease.

Upregulated Long Non-Coding RNA LL22NC03-N64E9.1 Promotes the Proliferation and Migration of Human Breast Cancer Cells by Silencing Kruppel-Like Factor 2 Expression.

INTRODUCTION: Recently, the significant regulatory effects of lncRNAs on the oncogenesis and growth of tumor have been demonstrated by an increasing number of research projects. A previous study showed that LL22NC03-N64E9.1 could promote the development of colorectal cancer, especially via enhanced cell proliferation. Similarly, this lncRNA should have comparable functions in breast cancer (BC), which requires in-depth investigation. Therefore, this study was designed to explore the correlation of LL22NC03-N64E9.1 with BC. METHODS: qRT-PCR was used to assess the relative expression of LL22NC03-N64E9.1 in BC tissues. Cell viability examination and colony formation experiments were performed to investigate the role of LL22NC03-N64E9.1 in BC cell's proliferation. Transwell assays were used to explore the effects of LL22NC03-N64E9.1 on BC cell's migration. RNA immunoprecipitation, chromosome immunoprecipitation assay and rescue experiments were performed to analyze the association of LL22NC03-N64E9.1 with target proteins and genes in BC cells. RESULTS: We identified that LL22NC03-N64E9.1 is an oncogene, upregulated in BC, which was verified in a cohort of 48 pairs of BC tissues. Based on the loss-of-function experiments, silencing LL22NC03-N64E9.1 expression significantly inhibited malignancy progression. In terms of the mechanism, LL22NC03-N64E9.1 acted on the enhancer of zeste homolog 2 (EZH2) by direct binding, which promoted BC cell growth. Furthermore, in the promoters of KLF2, the trimethylation of H3K27 could be regulated by LL22NC03-N64E9.1 as the mediator. CONCLUSION: Relying on the LL22NC03-N64E9.1/EZH2/KLF2 pathway, the lncRNA LL22NC03-N64E9.1 was significantly associated with BC development and could, therefore, be a potential therapeutic target to block BC growth.

Prognostic value of ephrin B receptors in breast cancer: An online survival analysis using the microarray data of 3,554 patients.

The roles of Ephrin B (EphB) receptors in cancer are relatively unknown as these receptors are associated with complex signaling pathways. A limited number of studies have investigated the association between EphB receptors and prognosis. Using the Kaplan-Meier plotter database, the present study investigated the associations between the mRNA expression levels of five EphB receptors and the outcomes of 3,554 patients with breast cancer who had been followed-up for 20 years. Hazard ratios (HR) and 95% confidence intervals (CI) were calculated to assess the relative risk of survival. The results demonstrated that high mRNA expression levels of EphB2 (HR, 0.74; 95% CI, 0.66-0.84; P=2.1×10-6), EphB4 (HR, 0.82; 95% CI, 0.72-0.93; P=0.0023) and EphB6 (HR, 0.69; 95% CI, 0.61-0.78; P=3×10-9) were significantly associated with improved survival, while a high mRNA expression level of EphB3 (HR, 1.14; 95% CI, 1.01-1.28; P=0.029) was associated with worse survival for patients with breast cancer. High expression levels of all EphB receptors, including EphB1 (HR, 1.4; 95% CI, 1.02-1.94; P=0.039), EphB2 (HR, 1.34; 95% CI, 1.07-1.67; P=0.011), EphB3 (HR, 1.39; 95% CI, 1.11-1.73, P=0.0038), EphB4 (HR, 1.33; 95% CI, 1.06-1.67; P=0.013) and EphB6 (HR, 1.32; 95% CI, 1.05-1.65; P=0.016), were associated with an increased risk of mortality in patients with lymph-node-positive breast cancer. High mRNA expression levels of EphB1 were not associated with survival for all patients with breast cancer (HR, 0.85; 95% CI, 0.72-1.01; P=0.058). The results of the present suggested that EphB receptors may be useful as prognostic biomarkers of breast cancer.

MicroRNA­126­3p inhibits the proliferation, migration, invasion, and angiogenesis of triple­negative breast cancer cells by targeting RGS3.

Triple­negative breast cancer (TNBC) is characterized by fast progression with high potential for metastasis, and poor prognosis. The dysregulation of microRNAs (miRNAs) occurring in the initiation or progression of cancers often leads to aberrant gene expression. The aim of the present study was to explore the function of miR­126 in TNBC cells. Expression levels of miR­126­3p were determined by quantitative real­time PCR. Then, the effects of miR­126­3p on migration, proliferation, invasion, and angiogenesis were assessed through in vitro experiments including Cell Counting Kit­8, colony formation, Transwell invasion and vasculogenic mimicry formation assays. One of the target genes for miR­126­3p predicted by TargetScan was confirmed by luciferase activity assay. Results indicated that miR­126­3p expression was reduced in TNBC cell lines. Functional assays revealed that miR­126­3p overexpression inhibited cell proliferation, migration, invasion, colony formation capacity and vasculogenesis by 1.2­, 1.8­, 2.3­, 2.0­ and 3.3­fold, respectively, compared to the miRNA­negative control group of MDA­MB­231 cells (P<0.001, respectively). In addition, the regulator of G­protein signaling 3 (RGS3) was hypothesized and validated as a direct target of miR­126­3p in TNBC. The proliferation, migration, invasion, colony formation capacity and vasculogenesis of MDA­MB­231 cells were significantly increased by 1.4­, 2.0­, 1.8­, 1.4­ and 3.2­fold, respectively, in cells transfected with pcDNA3.0­RGS3 compared to pcDNA3.0­negative control groups (P<0.001, respectively). The influence of miR­126­3p expression was reversed by RGS3 restoration. Collectively, the present study revealed that miR­126­3p plays a role as a tumor suppressor in regulating TNBC cell activities by targeting RGS3, indicating that the miR­126­3p/RGS3 axis may be a potential treatment target.

[Association between IL-6-572 C/G polymorphism and breast cancer susceptibility].

Objective To explore the associations between breast cancer and three single nucleotide polymorphisms (-597 G/A, -572 C/G and -174 G/C) of interleukin-6 (IL-6) gene promoter. Methods The study enrolled 136 breast cancer cases and 150 healthy female controls. The single nucleotide polymorphisms (SNPs) of IL-6 were identified by Sanger method of DNA sequencing, and serum IL-6 levels were measured with electrochemiluminescence immunoassay. Chi-squared test was utilized to compare frequency of a given SNP between the two groups, and t-test was employed to compare serum IL-6 levels in different groups. Results Compared with healthy controls, the serum IL-6 levels were significantly elevated in the patients with breast cancer. The genotype frequency of -572 C/G was significantly higher in breast cancer group than in the control group (OR=1.841, 95%CI: 1.115-3.040, P=0.017). There was no significant difference of serum IL-6 level in the three genotypes of -572 C/G, in both the breast cancer cases and the healthy controls. Conclusion The serum level of IL-6 significantly increases in the patients with breast cancer. The -572 C/G polymorphism and the serum level of IL-6 may play a role in the development of breast cancer.