Search

Rationale : Hemodynamic disturbed flow (DF) is associated with susceptibility to atherosclerosis. Endothelial Kruppel-Like Factor 4 (KLF4) is an important anti-inflammatory atheroprotective transcription factor that is suppressed in regions of DF. Objective : The plasticity of epigenomic KLF4 transcriptional regulation by flow-mediated DNA methylation was investigated in vitro and in arterial tissue. Methods and Results : To recapitulate dominant flow characteristics of atheroprotected and atherosusceptible arteries, human aortic endothelial cells were subjected to pulsatile undisturbed flow or oscillatory DF containing a flow-reversing phase. Differential CpG site methylation was measured by methylation-specific polymerase chain reaction, bisulfite pyrosequencing, and restriction enzyme-polymerase chain reaction. The methylation profiles of endothelium from disturbed and undisturbed flow sites of adult swine aortas were also investigated. In vitro, DF increased DNA methylation of CpG islands within the KLF4 promoter that significantly contributed to suppression of KLF4 transcription; the effects were mitigated by DNA methyltransferase (DNMT) inhibitors and knockdown of DNMT3A. Contributory mechanisms included DF-induced increase of DNMT3A protein (1.7-fold), DNMT3A enrichment (11-fold) on the KLF4 promoter, and competitive blocking of a myocyte enhancer factor-2 binding site in the KLF4 promoter near the transcription start site. DF also induced DNMT-sensitive propathological expression of downstream KLF4 transcription targets nitric oxide synthase 3, thrombomodulin, and monocyte chemoattractant protein-1. In support of the in vitro findings, swine aortic endothelium isolated from DF regions expressed significantly lower KLF4 and nitric oxide synthase 3, and bisulfite sequencing of KLF4 promoter identified a hypermethylated myocyte enhancer factor-2 binding site. Conclusions : Hemodynamics influence endothelial KLF4 expression through DNMT enrichment/myocyte enhancer factor-2 inhibition mechanisms of KLF4 promoter CpG methylation with regional consequences for atherosusceptibility.

Fetoplacental endothelial cells reside under physiological normoxic conditions (~2–8% O2) in vivo. Under such conditions, cells are believed to sense O2 changes primarily via hypoxia inducible factor 1 α(HIF1A). However, little is known regarding the role of HIF1A in fetoplacental endothelial function under physiological normoxia. We recently reported that physiological chronic normoxia (PCN; 20–25 day, 3% O2) enhanced FGF2- and VEGFA-stimulated proliferation and migration of human umbilical vein endothelial cells (HUVECs) via the MEK/ERK1/2 and PI3K/AKT1 pathways compared to standard cell culture normoxia (SCN; ambient O2: ~ 21% O2). Here, we investigated the action of HIF1A in regulating these cellular responses in HUVECs. HIF1A adenovirus infection in SCN-cells increased HIF1A protein expression, enhanced FGF2- and VEGFA-stimulated cell proliferation by 2.4 and 2.0 fold respectively, and promoted VEGFA-stimulated cell migration by 1.4 fold. HIF1A adenovirus infection in SCN-cells did not affect either basal or FGF2- and VEGFA-induced ERK1/2 activation, but it decreased basal AKT1 phosphorylation. Interestingly, HIF1A knockdown in PCN-cells via specific HIF1A siRNA transfection did not alter FGF2- and VEGFA-stimulated cell proliferation and migration, or ERK1/2 activation; however, it inhibited FGF2-induced AKT1 activation by ~ 50%. These data indicate that HIF1A differentially regulates cell proliferation and migration, and ERK1/2 and AKT1 activation in PCN- and SCN-HUVECs. These data also suggest that HIF1A critically regulates cell proliferation and migration in SCN-, but not in PCN-HUVECs.

BACKGROUND The GATA3 gene (GATA-binding protein 3) is one of the most frequently mutated genes in breast cancer. The objective of the current study was to determine the clinicopathologic characteristics of patients with breast cancer harboring GATA3 mutations. METHODS The authors examined the somatic mutation status of GATA3 and performed survival analysis in The Cancer Genome Atlas (TCGA) cohort (n = 934) and the Fudan University Shanghai Cancer Center (FUSCC) cohort (n = 308). Patient characteristics, including age; menopausal status; tumor laterality; tumor size; lymph node status; tumor grade; molecular subtypes; adjuvant radiotherapy, chemotherapy, and endocrine therapy; and prognosis, together with PIK3CA (phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha) and TP53 (tumor protein p53) mutation status, were collected. RESULTS GATA3 mutations were detected in 8.8% of patients (82 of 934 patients) in the TCGA cohort and 14.9% of patients (46 of 308 patients) in the FUSCC cohort. GATA3 mutations were found to be significantly associated with luminal-like breast cancer (P = .002 in the TCGA cohort and P

Triple-negative breast cancer (TNBC) patients have the worst prognosis and distant metastasis-free survival among all major subtypes of breast cancer. The poor clinical outlook is further exacerbated by a lack of effective targeted therapies for TNBC. Here we show that ectopic expression and therapeutic delivery of the secreted protein Tubulointerstitial nephritis antigen-like 1 (Tinagl1) suppresses TNBC progression and metastasis through direct binding to integrin α5β1, αvβ1, and epidermal growth factor receptor (EGFR), and subsequent simultaneous inhibition of focal adhesion kinase (FAK) and EGFR signaling pathways. Moreover, Tinagl1 protein level is associated with good prognosis and reversely correlates with FAK and EGFR activation status in TNBC. Our results suggest Tinagl1 as a candidate therapeutic agent for TNBC by dual inhibition of integrin/FAK and EGFR signaling pathways.

Purpose: This study sought to investigate the prognostic value of the autophagy marker microtubule–associated protein chain 3B (LC3B) in patients with residual tumors after neoadjuvant chemotherapy (NCT) for locally advanced breast cancer (LABC). Patients and Methods: The expression of LC3B in residual breast cancer cells was assessed by immunohistochemistry in surgical specimens from 229 patients diagnosed with histologically proven invasive breast cancer. All patients underwent NCT followed by mastectomy and were considered nonpathologic complete responders (non-pCR) after a pathologic evaluation. The prognostic value of various clinicopathologic factors was evaluated. Results: The LC3B density was similar between the peripheral and central area of the tumors (P = 0.328) but was significantly lower in the extratumoral area (P < 0.001 and P < 0.001, respectively). Furthermore, LC3B density, which correlated with Beclin-1 expression, Ki-67 index, and breast cancer subtype, served as an independent prognostic factor for both relapse-free survival (RFS; P = 0.012) and overall survival (OS; P = 0.008); the prognostic value of LC3B was most significant in triple-negative patients. Using a combination of LC3B expression and the status of residual involved lymph nodes, the patients were classified into four groups with different risks of relapse and death (P < 0.001 for RFS and P = 0.003 for OS). Conclusion: LC3B can be used as a prognostic marker in patients with non-pCR after NCT for breast cancer, which highlights the importance of autophagy in the biologic behavior of chemoresistant cancer cells. Furthermore, evaluating and targeting autophagy in the neoadjuvant setting may help prevent disease relapse in patients with non-pCR. Clin Cancer Res; 19(24); 6853–62. ©2013 AACR.

Purpose Triple-negative breast cancer (TNBC) is a highly diverse group of disease, and clinical outcome of patients with TNBC is highly variable. Due to the heterogeneity of TNBCs, there is limited molecular signature routinely used for predicting the risk of disease recurrence and benefit of adjuvant chemotherapy. Our study aims to develop and validate a RNA signature, integrating messenger RNAs (mRNAs) and long non-coding RNAs (lncRNAs) together, for TNBC patients to improve risk stratification and avoid unnecessary adjuvant therapy. Methods Using transcriptome microarrays, we analyzed 33 paired TNBC and adjacent normal breast tissues, and identified 1,644 mRNAs and 1,047 lncRNAs which were differentially expressed between tumors and normal tissues. We further determined the expression of these mRNAs and lncRNAs in an additional 134 TNBC samples using transcriptome microarrays, and confirmed their associations with patients' recurrence-free survival (RFS). Using the LASSO Cox regression model, we built an integrated mRNA-lncRNA signature incorporating seven mRNAs and three lncRNAs. Prognostic and predictive accuracy of the signature was tested in the training set of 167 TNBC patients and further validated in an independent validation set of 143 TNBC patients. Results In the training set, we identified 36 mRNAs and 32 lncRNAs which were tumor-specific and significantly associated with patients' RFS. Using the LASSO Cox regression model, an integrated mRNA-lncRNA signature based on seven mRNAs (CCR4, CTSB, ERO1L, HIF1A, IGFR1R, SRD5A1 and TFF1) and three lncRNAs (n381928, n333541 and TCONS_I2_00013109-XLOC_I2_007048) was developed in the training set and subsequently validated in the validation set. Patients were classified to the high-risk (high risk of recurrence) and low-risk (low risk of recurrence) groups according to their scores in the signature. In the training set, multivariate analysis showed that the predicted high-risk group had higher risk of developing recurrent disease within five years of surgery than the low-risk patients (hazard ratio [HR] = 6.12, 95% confidence interval [CI] 2.76-13.56, P Conclusion The integrated mRNA-lncRNA signature is a reliable tool for predicting disease recurrence and benefit of adjuvant chemotherapy in TNBC patients. If further validated in larger population, it could facilitate patient counseling and individualize treatment of TNBC. Citation Format: Yizhou Jiang, Yi-Rong Liu, Ke-Da Yu, Xin Hu, Xiao-En Xu, Ling Yao, Zhi-Ming Shao. Prognostic and predictive value of an integrated mRNA-lncRNA signature in triple-negative breast cancer: A comprehensive transcriptome analysis [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P6-08-28.

Ergonomics applications is now a highlight spot in various fields. To tackle the problem of unsustainable accumulation of ergonomics knowledge, the representation of ergonomics knowledge is of great importance for the ergonomics experts system to be developed. Based on the appropriate ontology technology, the general knowledge of ergonomics is reclassified to the man-machine-environment. By adopting the top-down method and Protégé, an ontology-based ergonomic framework is conducted to the ergonomics knowledge classification and representation.

Esophageal squamous cell carcinoma (ESCC) is a deadly malignancy with regard to mortality and prognosis, and the 5-year survival rate for all patients diagnosed with ESCC remains poor. A better understanding of the biological mechanisms of ESCC tumorigenesis and progression is of great importance to improve treatment of this disease. In this study, we demonstrated that the glutathione metabolism pathway is highly enriched in ESCC cells compared with normal esophageal epithelial cells in an in vivo mouse model. In addition, treatment with L-buthionine-sulfoximine (BSO) to deplete glutathione decreased the ESCC tumor burden in mice, thus demonstrating the critical role of glutathione metabolism in ESCC progression. BSO treatment also led to decreased cell proliferation and activation of cell apoptosis in ESCC. Finally, BSO treatment blocked NF-kB pathway activation in ESCC. Our study reveals a new pathway that regulates ESCC progression and suggests that inhibition of glutathione metabolism may be a potential strategy for ESCC treatment.

Assisted reproductive technology (ART) is widely used for the women with infertility conditions to achieve pregnancy. However, the adverse effects of ART may lead to poor perinatal and neonatal outcomes, e.g., preterm birth and low body weight. In this study, we investigated the inflammatory molecular factors and microRNA that might be involved in ART related preterm birth. We found the elevation of Toll-like 4 receptor (TLR4), activation of NFκB pathway and down-regulation of microRNA-146a (miR-146a), a negative regulator of NFκB, in the placenta of preterm birth and ART, indicating preterm birth and ART were associated with inflammation signaling activation. In vitro experiments demonstrated that miR-146a suppressed NFκB pathway and shifted the balance of cytokines in the cord blood toward a repertoire of pro-inflammatory outcomes by down-regulating IRAK1 and TRAF6. The pro-inflammatory cytokines IL-6, IFNγ and TNFα in the cord blood were highly expressed in the preterm and ART, while anti-inflammatory cytokine IL-10 was the lower in the preterm and ART. In summary, we firstly uncovered that TLR4/NFκB mediated inflammation signaling and miR-146a participated in ART-related preterm birth patients, which suggests that importance of TLR4/NFκB/miR-146a signaling in clinical interventions and biomarkers of ART-related perinatal or neonatal outcomes.

Long non-coding RNAs (lncRNAs) play an important role in gene regulation and are involving in diverse cellular processes. However, their roles in reprogramming of gene expression profiles during lineage commitment and maturation of mesenchymal stem cells (MSCs) remain poorly understood. In the current study, we characterize the expression of a lncRNA, HoxA-AS3, during the differentiation of MSCs. We showed that HoxA-AS3 is increased upon adipogenic induction of MSCs, while HoxA-AS3 remains unaltered during osteogenic induction. Silencing of HoxA-AS3 in MSCs resulted in decreased adipogenesis and expression of adipogenic markers, PPARG, CEBPA, FABP4 and ADIPOQ. Conversely, knockdown of HoxA-AS3 expression in MSCs exhibited an enhanced osteogenesis and osteogenic markers expression, including RUNX2, SP7, COL1A1, IBSP, BGLAP and SPP1. Mechanistically, HoxA-AS3 interacts with Enhancer Of Zeste 2 (EZH2) and is required for H3 lysine-27 trimethylation (H3K27me3) of key osteogenic transcription factor Runx2. Our data reveal that HoxA-AS3 acts as an epigenetic switch that determines the lineage specification of MSC.

The value of sentinel lymph node biopsy (SLNB) in post-neoadjuvant chemotherapy (NCT) patients is still controversial. We aimed to identify predictors and construct a nomogram for predicting the pathologically complete response (pCR) of axillary lymph nodes (ALNs) after NCT in node positive breast cancer patients. In total, 426 patients with pathologically proven ALN metastasis before NCT were enrolled, randomized 1:1 and divided into a training set and a validation set. We developed a nomogram based on independent predictors for ALN pCR identified by multivariate logistic regression as well as clinical significant predictors. The multivariate logistic regression analysis showed that hormone receptor (HR) status, human epidermal growth factor 2 (HER2) status and Ki67 index were independent predictors. The nomogram was thereby constructed by those independent predictors as well as tumor size and NCT regimens. The areas under the receiver operating characteristic curve of the training set and the validation set were 0.804 and 0.749, respectively. We constructed a nomogram for predicting ALN pCR in patients who received NCT. Our nomogram can improve risk stratification, accurately predict post-NCT ALN status and avoid unnecessary ALN dissection.

Background Unlike arteries, in which regionally distinct hemodynamics are associated with phenotypic heterogeneity, the relationships between endocardial endothelial cell phenotype and intraventricular flow remain largely unexplored. We investigated regional differences in left ventricular wall shear stress and their association with endocardial endothelial cell gene expression. Methods and Results Local wall shear stress was calculated from 4‐dimensional flow magnetic resonance imaging in 3 distinct regions of human (n=8) and pig (n=5) left ventricle: base, adjacent to the outflow tract; midventricle; and apex. In both species, wall shear stress values were significantly lower in the apex and midventricle relative to the base; oscillatory shear index was elevated in the apex. RNA sequencing of the endocardial endothelial cell transcriptome in pig left ventricle (n=8) at a false discovery rate ≤10% identified 1051 genes differentially expressed between the base and the apex and 327 between the base and the midventricle; no differentially expressed genes were detected at this false discovery rate between the apex and the midventricle. Enrichment analyses identified apical upregulation of genes associated with translation initiation including mammalian target of rapamycin, and eukaryotic initiation factor 2 signaling. Genes of mitochondrial dysfunction and oxidative phosphorylation were also consistently upregulated in the left ventricular apex, as were tissue factor pathway inhibitor (mean 50‐fold) and prostacyclin synthase (5‐fold)—genes prominently associated with antithrombotic protection. Conclusions We report the first spatiotemporal measurements of wall shear stress within the left ventricle and linked regional hemodynamics to heterogeneity in ventricular endothelial gene expression, most notably to translation initiation and anticoagulation properties in the left ventricular apex, in which oscillatory shear index is increased and wall shear stress is decreased.

Ping Zhou,1–3 Yi-Zhou Jiang,4 Xin Hu,4 Wei Sun,4 Yi-Rong Liu,4 Fang Liu,5 Rong-Cheng Luo,1,* Zhi-Ming Shao4,* 1Department of Oncology, TCM-Integrated Cancer Center of Southern Medical University, Southern Medical University, Guangzhou, Guangdong, 2Department of Breast Surgery, The Third Hospital of Nanchang, 3Jiangxi Provincial Key Laboratory for Breast Diseases, Jiangxi, 4Department ofBreast Surgery, Fudan University Shanghai Cancer Center/Cancer Institute, Shanghai, 5Department ofPathophysiology, Foshan University, Guangdong, People’s Republic ofChina *These authors contributed equally tothis work Objective: The aim of this study was to describe the clinical features and outcomes of Chinese patients with human epidermal growth factor receptor 2 (HER2)-positive breast cancer.Method: The clinical data and survival statuses of 732 patients with operable HER2-positive breast cancer who were treated at the Department of Breast Surgery of the Shanghai Cancer Center from January 1, 2007, to December 31, 2011, were collected. The patients were divided into two groups according to treatment with and without trastuzumab. Disease-free survival (DFS) and overall survival were calculated using the Kaplan–Meier method and log-rank test. The associations of the patient characteristics with prognosis were analyzed via Cox regression.Results: A total of 732 women with HER2-positive breast cancer were included in this study, among whom 258 (35.2%) received trastuzumab. The median follow-up duration was 41months. By the end of the follow-up period, 86 (12%) women experienced local recurrence or metastasis. Patients who received both anti-HER2 therapy and chemotherapy exhibited a longer DFS than those who received chemotherapy alone (P=0.001). Tumor size, lymph node status, and family history of breast cancer were associated with median DFS, and tumor size, lymph node status, clinical stage, age, and body mass index were associated with median overall survival. Patients who received both neoadjuvant chemotherapy and trastuzumab exhibited a higher rate of pathological complete remission. In the neoadjuvant group, the patients who received both anti-HER2 therapy and chemotherapy exhibited a longer DFS than those who received chemotherapy alone (P=0.049).Conclusion: Significant clinical features were observed in the Chinese patients with HER2-positive breast cancer. Furthermore, targeted anti-HER2 therapy may improve the prognosis of these patients. Keywords: breast cancer, trastuzumab, HER2, disease-free survival, chemotherapy

// Wei Sun 1, 2, 3, * , Yi-Zhou Jiang 1, 2, 3, * , Yi-Rong Liu 1, 2, 3, * , Ding Ma 1, 2, 3 , Zhi-Ming Shao 1, 2, 3, 4 1 Department of Breast Surgery, Fudan University Shanghai Cancer Center, Shanghai, P.R. China 2 Cancer Institute, Fudan University Shanghai Cancer Center, Shanghai, P.R. China 3 Department of Oncology, Shanghai Medical College, Fudan University, Shanghai, P.R. China 4 Institutes of Biomedical Sciences, Fudan University, Shanghai, P.R. China * These authors contributed equally to this work Correspondence to: Zhi-Ming Shao, e-mail: zhimingshao@yahoo.com Keywords: nomogram, luminal breast cancer, overall survival, breast cancer-specific survival Received: December 02, 2015 Accepted: February 13, 2016 Published: March 07, 2016 ABSTRACT Luminal breast cancer constitutes a group of highly heterogeneous diseases with a sustained high risk of late recurrence. We aimed to develop comprehensive and practical nomograms to better estimate the long-term survival of luminal breast cancer. Patients with luminal breast cancer diagnosed between 1990 and 2006 were retrieved from the Surveillance, Epidemiology, and End Results (SEER) database and randomly divided into the training ( n = 87,867) and validation ( n = 88,215) cohorts. The cumulative incidence function (CIF) and a competing-risks model were used to estimate the probability of breast cancer-specific survival (BCSS) and death from other causes. We integrated significant prognostic factors to build nomograms and subjected the nomograms to bootstrap internal validation and to external validation. We screened 176,082 luminal breast cancer cases. The 5- and 10-year probabilities of overall death were 0.089 and 0.202, respectively. The 5- and 10-year probabilities of breast cancer-specific mortality (BCSM) were 0.053 and 0.112, respectively. Nine independent prognostic factors for both OS and BCSS were integrated to construct the nomograms. The calibration curves for the probabilities of 5- and 10-year OS and BCSS showed excellent agreement between the nomogram prediction and actual observation. The C-indexes of the nomograms were high in both internal validation (0.732 for OS and 0.800 for BCSS) and external validation (0.731 for OS and 0.794 for BCSS). We established nomograms that accurately predict OS and BCSS for patients with luminal breast cancer. The nomograms can identify patients with higher risk of late overall mortality and BCSM, helping physicians in facilitating individualized treatment.

While recognized as a generally aggressive disease, triple-negative breast cancer (TNBC) is highly diverse in different patients with variable outcomes. In this prospective observational study, we aimed to develop an RNA signature of TNBC patients to improve risk stratification and optimize the choice of adjuvant therapy. Transcriptome microarrays for 33 paired TNBC and adjacent normal breast tissue revealed tumor-specific mRNAs and long noncoding RNAs (lncRNA) that were associated with recurrence-free survival. Using the Cox regression model, we developed an integrated mRNA-lncRNA signature based on the mRNA species for FCGR1A, RSAD2, CHRDL1, and the lncRNA species for HIF1A-AS2 and AK124454. The prognostic and predictive accuracy of this signature was evaluated in a training set of 137 TNBC patients and then validated in a second independent set of 138 TNBC patients. In addition, we enrolled 82 TNBC patients who underwent taxane-based neoadjuvant chemotherapy (NCT) to further verify the predictive value of the signature. In both the training and validation sets, the integrated signature had better prognostic value than clinicopathologic parameters. We also confirmed the interaction between the administration of taxane-based NCT and different risk groups. In the NCT cohort, patients in the low-risk group were more likely to achieve pathologic complete remission after taxane-based NCT (P = 0.014). Functionally, we showed that HIF1A-AS2 and AK124454 promoted cell proliferation and invasion in TNBC cells and contributed there to paclitaxel resistance. Overall, our results established an integrated mRNA-lncRNA signature as a reliable tool to predict tumor recurrence and the benefit of taxane chemotherapy in TNBC, warranting further investigation in larger populations to help frame individualized treatments for TNBC patients. Cancer Res; 76(8); 2105–14. ©2016 AACR.

Background: The effect of zoledronic acid in breast cancer adjuvant therapy concerning improvement of patient survival has yet to be confirmed. We performed a meta-analysis of published and unpublished randomized controlled trials with the aim of accurate evaluation between clinical outcome and the association of the addition of zoledronic acid to adjuvant therapy. Methods: We searched Pubmed (from 1966 to present) and online abstracts from the proceeding Annual Meetings of the American Society of Clinical Oncology (ASCO) (years 1992–2010) and online abstracts from San Antonio Breast Cancer Symposium (years 2004–2010). A total of five eligible studies including 3676 subjects and 3678 controls met our search criteria and were evaluated. Random and fixed-effects meta-analytical models were used where indicated, and between-study heterogeneity was assessed. The primary study endpoints were the DFS. Secondary endpoints were OS, distant or loco-regional recurrence free survival and bone metastasis free survival. Results: Compared with the control arm, adjuvant breast cancer treatment with zoledronic acid did not significantly improve overall survival (OS), disease free survival (DFS), bone metastasis free survival, distant and locoregional recurrence free survival. However, in the postmenopausal subgroup, the addition of zoledronic acid to standard therapy could significantly improve DFS (RR=0.763, 95%CI 0.658−0.884, p Discussion: Adjuvant zoledronic acid may potentially improve the prognosis of postmenopausal patients. Additional studies are needed to evaluate the value of adjuvant treatment of zoledronic acid in premenopausal couterparts, differing disease stages, and various pathological types of breast cancer. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P3-16-10.

Triple-negative breast cancer (TNBC), with a lack expression of ER, PR, and HER-2 as potential treatment targets, is insensitive to the hormonal and trastuzumab therapies. Thus, local and regional treatment, such as radiotherapy is placed with more emphasis when treating TNBC patients. In the current study, expression profiles of mRNAs and long noncoding RNAs (lncRNAs) in TNBC patients, who received radiotherapy in our institute, was investigated to develop radiation-related gene signatures to facilitate individualized TNBC treatment. Specifically, 44 TNBC samples including 14 recurrences and 30 non-recurrences after radiotherapy were analyzed by using transcriptome microarrays. A total of 626 mRNAs were identified as differentially expressed (DEGs) between recurrent patients and non-recurrent patients (|fold change| > 1.5, p < 0.05). GeneOntology analysis showed that these DEGs were enriched in immune response, mitotic cell cycle, inflammatory response, cell adhesion as well as extracellular matrix organization, which indicated that these biological processes may exert significant impacts on the therapeutic effects of radiation. In addition, a total of 150 lncRNAs were identified as differentially expressed between recurrent patients and non-recurrent patients (|fold change| > 1.5, p < 0.05). The differentially expressed mRNAs and lncRNAs were confirmed by quantitative real-time PCR. Cox regression model was further applied to explore potential lncRNAs involved in certain signaling pathways such as JAK/STAT, EGRF/ATK/PI3K are highly co-expressed with mRNAs in these signaling pathways. Overall, our results revealed a panel of mRNAs and lncRNAs with significant impacts on evaluating the TNBC radio-therapeutic effects as well predicting the possibility of recurrence. Citation Format: Xingxing CHEN, Zhaozhi YANG, Yizhou JIANG, Jinli MA, Xiaoli YU, Zhimin SHAO, Xiaomao GUO. Comprehensive transcriptome analysis identifies integrated mRNA-lncRNA signature of triple-negative breast cancer treated with radiotherapy [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 3198.

Long non-coding RNAs (lncRNAs) play an important role in gene regulation and are involving in diverse cellular processes. However, their roles in reprogramming of gene expression profiles during lineage commitment and maturation of mesenchymal stem cells (MSCs) remain poorly understood. In the current study, we characterize the expression of a lncRNA, HoxA-AS3, during the differentiation of MSCs. We showed that HoxA-AS3 is increased upon adipogenic induction of MSCs, while HoxA-AS3 remains unaltered during osteogenic induction. Silencing of HoxA-AS3 in MSCs resulted in decreased adipogenesis and expression of adipogenic markers, PPARG, CEBPA, FABP4 and ADIPOQ. Conversely, knockdown of HoxA-AS3 expression in MSCs exhibited an enhanced osteogenesis and osteogenic markers expression, including RUNX2, SP7, COL1A1, IBSP, BGLAP and SPP1. Mechanistically, HoxA-AS3 interacts with Enhancer Of Zeste 2 (EZH2) and is required for H3 lysine-27 trimethylation (H3K27me3) of key osteogenic transcription factor Runx2. Our data reveal that HoxA-AS3 acts as an epigenetic switch that determines the lineage specification of MSC.

The aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor, mediates many biological processes, including fetal development. In this study, we examined AhR protein expression in human placentas from normal (N) and severe preeclamptic (sPE) pregnancies, as well as human fetal tissues from the second trimester of pregnancy, using immunohistochemistry and/or Western blot analysis. In the placentas, the AhR immunoreactivity was present primarily in syncytiotrophoblasts. The AhR staining was also seen in endothelium of large blood vessels in villi and endothelium of umbilical cord arteries and veins. No difference in AhR protein levels was found between N and sPE placentas. In fetal tissues, the AhR immunoreactivity was localized in lung, kidney, esophagus, pancreas, liver, testicle, thymus gland, retina, and choroid, mainly in epithelial cells, whereas it was absent in heart, brain, sclera, and thoracic aorta. These findings suggest that the AhR plays a critical role in syncytiotrophoblasts of human placentas and epithelium of many fetal organs. These data also imply that human placentas and those fetal organs with high AhR expression (e.g., lung, kidney, liver, pancreas, and thymus gland) during fetal development are highly susceptible to environmental toxicants such as dioxin. (J Histochem Cytochem 58:679–685, 2010)

Placental development occurs under a low oxygen (2-8% O(2)) environment, which is critical for placental development and angiogenesis. In this study, we examined if hypoxia affected fibroblast growth factor-2 (FGF2)- and vascular endothelial growth factor (VEGF)-stimulated cell proliferation via the mitogen-activated protein kinase kinase 1/2 (MEK1/2)/extracellular signal-regulated kinases 1/2 (ERK1/2) and phosphatidylinositol-3 kinase (PI3K)/v-akt murine thymomaviral oncogene homologue (AKT1) pathways in human placental artery endothelial (HPAE) cells. We observed that under normoxia (approximately 20% O(2)), FGF2 and VEGF dose-dependently stimulated cell proliferation. Hypoxia (3% O(2)) significantly promoted FGF2- and VEGF-stimulated cell proliferation as compared to normoxia. Under both normoxia and hypoxia, FGF2 rapidly induced ERK1/2 and AKT1 phosphorylation, while VEGF-induced ERK1/2, but not AKT1 phosphorylation. However, hypoxia did not significantly alter FGF2- and VEGF-induced ERK1/2 and AKT1 phosphorylation as compared to normoxia. PD98059 (a MEK1/2 inhibitor) at 20microM and LY294002 (a PI3K inhibitor) at 5microM attenuated FGF2- and VEGF-induced phosphorylation of ERK1/2 and AKT1, respectively. PD98059, even at doses that drastically inhibited FGF2-induced ERK1/2 phosphorylation (20microM) and caused cell loss (40microM), did not affect FGF2-stimulated cell proliferation, which was confirmed by U0126 (another potent MEK1/2 inhibitor). PD98059, however, dose-dependently inhibited VEGF-stimulated cell proliferation. Conversely, LY294002 dose-dependently inhibited FGF2-, but not VEGF-stimulated cell proliferation. These data suggest that in the MEK1/2/ERK1/2 and PI3K/AKT1 pathways differentially mediate FGF2- and VEGF-stimulated HPAE cell proliferation. These results also indicate that hypoxia promotes FGF2- and VEGF-stimulated cell proliferation without further activation of the PI3K/AKT1 and MEK1/2/ERK1/2, respectively.

Background The clinical significance of progesterone receptor (PgR) expression in estrogen receptor-negative (ER–) breast cancer is controversial. Herein, we systemically investigate the clinicopathologic features, molecular essence, and endocrine responsiveness of ER−/PgR+/HER2− phenotype. Methods Four study cohorts were included. The first and second cohorts were from the Surveillance, Epidemiology, and End Results database (n = 67,932) and Fudan University Shanghai Cancer Center (n = 2,338), respectively, for clinicopathologic and survival analysis. The third and fourth cohorts were from two independent publicly available microarray datasets including 837 operable cases and 483 cases undergoing neoadjuvant chemotherapy, respectively, for clinicopathologic and gene-expression analysis. Characterized genes defining subgroups within the ER–/PgR+/HER2– phenotype were determined and further validated. Results Clinicopathologic features and survival outcomes of the ER–/PgR+ phenotype fell in between the ER+/PgR+ and ER−/PgR− phenotypes, but were more similar to ER−/PgR−. Among the ER−/PgR+ phenotype, 30 % (95 % confidence interval [CI] 17–42 %, pooled by a fixed-effects method) were luminal-like and 59 % (95 % CI 45–72 %, pooled by a fixed-effects method) were basal-like. We further refined the characterized genes for subtypes within the ER−/PgR+ phenotype and developed an immunohistochemistry-based method that could determine the molecular essence of ER−/PgR+ using three markers, TFF1, CK5, and EGFR. Either PAM50-defined or immunohistochemistry-defined basal-like ER−/PgR+ cases have a lower endocrine therapy sensitivity score compared with luminal-like ER−/PgR+ cases (P

Triple-negative breast cancer (TNBC) is a heterogeneous disease with highest loco-regional recurrence among breast cancer subtypes. Radiotherapy is indispensable for TNBC loco-regional control. However, intrinsic radiosensitivity differences exist in TNBC patients and RT is still prescribed mainly based on conventional clinicopathologic features of patients without considering the differences. The purpose of the present study is to develop and validate a TNBC radiosensitive gene signature (RSGS) and to guide therapeutic decisions. In this study, we compared transcriptome profiles of 12 locally recurrent TNBCs to 20 non-locally recurrent TNBCs treated with surgery radio-chemotherapy and developed a seven-gene RSGS and a simplified three-gene RSGS by using pathway analysis, univariate Cox proportional hazards regression model and rank-based linear algorithm. They were validated by using transcriptome profiles of 166 TNBC patients. Two gene signatures specifically identified a radiosensitive population that had an improved recurrence-free survival in patients treated with surgery radio-chemotherapy (Radiosensitive patients vs radioresistant patients, for seven-gene RSGS: P = 0.024, HR = 0.35, 95 %CI 0.14–0.87 and for three-gene RSGS: P = 0.035, HR = 0.38, 95 %CI 0.15–0.94). In contrast, there was no significant difference in outcome between predicted radiosensitive and radioresistant patients that treated with other treatment modality. RSGSs provide a useful tool for identification of radiosensitive/radioresistant TNBC patients and they could lead to a better selection of patients for RT protocols.

Background Atherosclerosis is a heterogeneously distributed disease of arteries in which the endothelium plays an important central role. Spatial transcriptome profiling of endothelium in pre-lesional arteries has demonstrated differential phenotypes primed for athero-susceptibility at hemodynamic sites associated with disturbed blood flow. DNA methylation is a powerful epigenetic regulator of endothelial transcription recently associated with flow characteristics. We investigated differential DNA methylation in flow region-specific aortic endothelial cells in vivo in adult domestic male and female swine. Results Genome-wide DNA methylation was profiled in endothelial cells (EC) isolated from two robust locations of differing patho-susceptibility: − an athero-susceptible site located at the inner curvature of the aortic arch (AA) and an athero-protected region in the descending thoracic (DT) aorta. Complete methylated DNA immunoprecipitation sequencing (MeDIP-seq) identified over 5500 endothelial differentially methylated regions (DMRs). DMR density was significantly enriched in exons and 5’UTR sequences of annotated genes, 60 of which are linked to cardiovascular disease. The set of DMR-associated genes was enriched in transcriptional regulation, pattern specification HOX loci, oxidative stress and the ER stress adaptive pathway, all categories linked to athero-susceptible endothelium. Examination of the relationship between DMR and mRNA in HOXA genes demonstrated a significant inverse relationship between CpG island promoter methylation and gene expression. Methylation-specific PCR (MSP) confirmed differential CpG methylation of HOXA genes, the ER stress gene ATF4, inflammatory regulator microRNA-10a and ARHGAP25 that encodes a negative regulator of Rho GTPases involved in cytoskeleton remodeling. Gender-specific DMRs associated with ciliogenesis that may be linked to defects in cilia development were also identified in AA DMRs. Conclusions An endothelial methylome analysis identifies epigenetic DMR characteristics associated with transcriptional regulation in regions of atherosusceptibility in swine aorta in vivo. The data represent the first methylome blueprint for spatio-temporal analyses of lesion susceptibility predisposing to endothelial dysfunction in complex flow environments in vivo. Electronic supplementary material The online version of this article (doi:10.1186/s12864-015-1656-4) contains supplementary material, which is available to authorized users.

Guo-Wei Qian,1,* Xiao-Jian Ni,1,* Zheng Wang,2 Yi-Zhou Jiang,1 Ke-Da Yu,1 Zhi-Ming Shao1 1Department of Breast Surgery, 2Department of Radiation Oncology, Shanghai Cancer Center and Cancer Institute, Fudan University, Shanghai, People’s Republic of China *These authors contributed equally tothis work Background: Although it has been previously reported that radiotherapy (RT) effectively reduced the incidence of local recurrence of ductal carcinoma in situ (DCIS) following breast-conserving surgery (BCS), little is known about the effect of RT on survival of patients with locally excised DCIS. Patients and methods: Using Surveillance, Epidemiology, and End Results registry data, we selected 56,968female DCIS patients treated with BCS between 1998and 2007. Overall survival (OS) and breast cancer-specific survival (BCSS) were compared among patients who received RT or no RT using the Kaplan–Meier methods and Cox proportional hazards regression models. Results: Median follow-up was 91months. In the multivariable model, patients receiving postoperative RT had better OS than those undergoing BCS alone (hazard ratio [HR] 0.59, 95% confidence interval [CI] 0.53–0.67, P

Objective: One of the first clinically detectable changes in the vasculature during atherogenesis is the accumulation of cholesterol within the vessel wall. Hypercholesterolemia is characterized by dysfunctional endothelial-dependent vessel relaxation and impaired NOS3 function. Since DNA methylation at gene promoter regions strongly suppresses gene expression, we postulated that high-fat/high-cholesterol diet suppresses endothelial NOS3 through promoter DNA methylation. Methods: Domestic male pigs were fed control diet (CD) or isocaloric high fat and high cholesterol diet (HC; 12% fat and 1.5% cholesterol) for 2, 4, 8 or 12 weeks prior to tissue collection. Furthermore, to determine the effects of risk factor withdrawal, an additional group of swine received HC for 12 weeks and then CD for 8 weeks; a control group received HC continuously for 20 weeks. Endothelial cells were harvested from common carotid aorta. In parallel in vitro studies, cultured human aortic endothelial cells (HAEC) were treated with human LDL, GW3956 (LXR agonist) and RG108 (DNA methyltransferase [DNMT] inhibitor). In cells from both sources, DNA methylation at the NOS3 promoter was measured using methylation specific pyro sequencing, and endothelial gene expression was measured using RT PCR. Results: HC diet increased plasma cholesterol level from 75 mg/dl on CD to a plateau of about 540 mg/dl within 2 weeks. Endothelial NOS3 expression was significantly reduced (71±9 % of CD) after 4 weeks of HC, a level sustained at subsequent time points. Withdrawal of HC for 8 weeks did not recover NOS3 expression. After 12-week HC, the NOS3 promoter was hypermethylated. Withdrawal of HC did not reverse NOS3 promoter methylation. In vitro treatment of HAEC with human LDL (200 mg/dl total cholesterol) or GW3956 (5μM) suppressed NOS3 mRNA to 50% and 30% respectively, suggesting that LXR/RXR is involved in suppression of NOS3. Nitric oxide production was consistently suppressed by GW3959. Both could be reversed through inhibition of DNMTs by RG108. Conclusions: DNA methylation and LXR/RXR pathway can mediate the HC-suppression of endothelial NOS3. The study identifies novel pharmaceutical targets in treating endothelial dysfunction. Crosstalk between these pathways is under investigation.

// Ke-Da Yu 1 , Yi-Zhou Jiang 1 , Zhi-Ming Shao 1 1 Department of Breast Surgery, Cancer Center and Cancer Institute, Shanghai Medical College, Fudan University, Shanghai, P.R. China Correspondence to: Ke-Da Yu, e-mail: yukeda@163.com Keywords: breast cancer, metastatic potential, lymph node Received: February 15, 2015 Accepted: March 24, 2015 Published: April 10, 2015 ABSTRACT Purpose: Poor prognosis associated with metastasis in breast cancer patients highlights the critical need to develop an effective evaluation model for metastatic potential (MP). We hypothesized that MP could be also indicated by primary tumor size and involved lymph nodes (LNs). Methods: The expected number of involved LNs is defined as tumor size (cm) divided by 1.5. The effect of the surrogate for MP (defined as difference between the number of observed and expected involved LNs) on breast cancer-specific survival (BCSS) was investigated in the first cohort from SEER ( n = 108,814). Validation was performed in another SEER cohort ( n = 50,414) and a third cohort ( n = 3,755). Results: MP is an independent predictor for BCSS in the overall population [hazard ratio (HR) for high MP: 2.92; 95% confidence interval (CI): 2.80–3.03] and in subgroups. The effect of surrogate for MP on survival was independent to intrinsic subtype, with adjusted HRs of 3.46 (95%CI, 2.02–5.93), 2.30 (95%CI, 1.64–3.24), 4.05 (95%CI, 2.85–5.76), and 1.45 (95%CI, 1.04–2.03) in luminal-A, luminal-B, triple-negative, and HER2-positive subtypes, respectively. Conclusion: Difference between the observed and expected number of involved LNs serves as an indicator for MP, which is independent to intrinsic subtype and could predict survival. Our findings need further validation.

Chinese breast cancer patients living in the United States (US) can experience different disease patterns than Caucasians, which might allow for predicting the future epidemiology of breast cancer in China. We aimed to compare the clinicopathologic characteristics and outcomes of Caucasian and Chinese female breast cancer patients residing in the US. The study cohort consisted of 3868 Chinese and 208621 Caucasian women (diagnosed from 1990 to 2009) in the US Surveillance, Epidemiology, and End Results (SEER) database. Compared with the Caucasian patients, the US-residing Chinese patients had a younger age at diagnosis and a higher family income, remained married longer, and more frequently lived in metropolitan areas. Other tumor characteristics were similarly distributed between the two races. Compared with the Caucasians, the Chinese patients had a significantly improved overall survival (OS) but similar breast cancer-specific survival (BCSS). Our analysis suggested that US-residing Chinese patients had significant differences in age, family income, marital status and area of residence, compared with their Caucasian counterparts. No significant disparities were noted in BCSS between the two races, whereas the Chinese patients had a significantly better OS. These findings warrant further investigation and should be considered in the screening and treatment of breast cancer.

Mesenchymal Stem Cells (MSC) can be obtained from various tissues and differentiate into many different lineages, including osteoblasts, adipocytes, chondrocytes, cardiomyocytes, hepatocytes and neural cells both in vivo and in vitro. However, the ability of MSC to differentiate into specific lineages seems to be restricted and requires a deeper understanding of the genetic and epigenetic mechanisms. Epigenetic mechanism refers to a process that regulates heritable alterations in gene expression without changing the DNA sequence. SWI/SNF (SWItch/Sucrose Non-Fermentable), a chromatin-remodeling complex serves as an ideal intervention point for lineage manipulation of MSC. In this review, we discuss the importance of SWI/SNF chromatin remodeling complex in regulating the fate determination of MSC. We propose that selectively manipulation of subunits of SWI/SNF will enhance the lineagespecific differentiation of MSC and improve therapeutic application of MSC.

This paper analyzes the classification, performance and application of ship fireproof and heat insulating materials, and describes the test standard and performance evaluation criteria of the non-combustibility, low flame-spread characteristics and smoke and toxicity of marine fireproof materials in detail. So the paper has certain reference value and guidance significance for the selection of heat insulating materials with fire divisions and the use of flammable materials on board in accordance with requirements.

Stem cell-mediated therapy is a promising strategy for treating pancreatic diseases such as Type-1 diabetes (T1D) and pancreatic cancers. Although islet transplantation has been reported to be an effective diabetes therapy, its worldwide application is extremely limited due to the shortage of donor islets and immune rejection problems. Stem cell-based approach for islet neogenesis in vivo could provide a promising alternative source of islets for treating diabetes. On the other hand, targeting the cancer stem cells could be very effective for the treatment of pancreatic cancers. In this review, we focused on the present progress in the field of adult pancreatic stem cells, stem cell-mediated strategies for treating T1D, and pancreatic cancer stem cells, while discussing of the possible challenges involved in them.

Purpose Neoadjuvant chemotherapy (NCT) is a standard treatment option for locally advanced breast cancer. However, the lack of an efficient method to predict treatment response and patient prognosis hampers the clinical evaluation of patient eligibility for NCT. An elevated lymphocyte-to-monocyte ratio (LMR) has been reported to be associated with a favorable prognosis for certain hematologic malignancies and for nasopharyngeal carcinoma; however, this association has not been investigated in breast cancer. The purpose of this study was to evaluate whether pre-NCT LMR analysis could predict the prognosis of patients with locally advanced breast cancer. Methods A retrospective cohort of 542 locally advanced breast cancer patients (T3/T4 and/or N2/N3 disease) receiving NCT followed by radical surgery was recruited between May 2002 and August 2011 at the Fudan University Shanghai Cancer Center. Counts for pre-NCT peripheral absolute lymphocytes and monocytes were obtained and used to calculate the LMR. Results Univariate and multivariate analysis revealed that higher LMR levels (≥4.25) were significantly associated with favorable DFS (P = 0.009 and P = 0.011, respectively). Additionally, univariate analysis revealed that a higher lymphocyte count (≥1.5×109/L) showed borderline significance for improved DFS (P = 0.054), while a lower monocyte count (

Few studies have been undertaken to evaluate the prognostic value of age at diagnosis for determining breast cancer survival in a large population.Using the U.S. National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) database consisting of 18 population-based cancer registries, this study identified 331,969 female patients with a diagnosis of breast cancer from 1 January 1990, to 31 December 31. Breast cancer-specific mortality (BCSM) was compared among patients in different age groups using Kaplan-Meier plots. The Cox proportional hazards model was used for multivariate analysis.In the multivariate analysis, the hazard ratios (HRs) of BCSM in the different age groups formed a U-shaped curve, with patients younger than 30 years and patients older than 79 years experiencing the worst survival rates (HR, 1.19; 95 % confidence interval [CI], 1.06-1.33; P = 0.003 and HR, 2.16; 95 % CI, 2.05-2.27; P0.001, with age 50-59 years as the reference, respectively). When the interaction between age at diagnosis and hormone receptor (HoR) status for prediction of BCSM was further analyzed, the findings showed that in the HoR-positive group, patients younger than 30 years and patients older than 79 years had the worst survival rates (HR, 1.52; 95 % CI, 1.30-1.76; P0.001 and HR, 2.07; 95 % CI, 1.94-2.20; P0.001, respectively), whereas patients ages 40 to 49 years had the best survival rate (HR, 0.93; 95 % CI, 0.89-0.98; P = 0.005). This pattern, however, was different in the HoR-negative group. Patients younger than 60 years had nearly the same BCSM (P = 0.356, 0.199, and 0.036 for ages30 years, 30-39 years, and 40-49 years, respectively), with BCSM starting to increase with age only for patients older than 60 years and peaking for patients older than 79 years (HR, 2.39; 95 % CI, 2.20-2.59; P0.001).The study findings show different patterns in the prognostic value of age for determining BCSM, depending on the HoR status. These data underscore the importance of age-specific studies for different HoR groups to individualize treatment and improve outcomes for breast cancer patients.

We investigated the loss of somatic mutations in TP53 and PIK3CA in breast cancer tissue after neoadjuvant chemotherapy (NCT) and the clinical relevance of the observed mutation profiles. Samples were derived from three cohorts: Cohort 1 consisting of 206 patients undergoing NCT with matched pre- and postchemotherapy tumor tissues; Cohort 2 consisting of 158 additional patients undergoing NCT; and Cohort 3, consisting of 81 patients undergoing chemotherapy with prechemotherapy tumor tissues. In the first cohort, somatic mutations in TP53 or PIK3CA were identified in 24.8% of the pre-NCT tumor samples but in only 12.1% of the post-NCT tumor samples (P < 0.001). Patients with initial TP53 and PIK3CA mutations who became negative for the mutations after NCT had a higher Miller–Payne score (P = 0.008), improved disease-free survival, and improved overall survival than those with no change or the opposite change. The association of loss of mutations in TP53 and PIK3CA and improved survival was successfully validated in the second cohort. In addition, 28.4% of the tumors showed intratumoral heterogeneity of somatic mutations in TP53 or PIK3CA, whereas 71.6% were homogeneous, either with or without the mutations. Our data reveal the novel concept that chemotherapy may reduce mutation frequency in patients with breast cancer. Furthermore, the loss of somatic mutations in TP53 and PIK3CA may be translated to biomarkers for prognosis via further verification, which may optimize the choice of sequential therapy and improve patient survival. Cancer Res; 74(13); 3399–407. ©2014 AACR.

Background Reduced overall survival has been observed in patients with left-sided versus right-sided breast cancer due to cardiac toxicity after radiotherapy. However, the effect of laterality and primary tumor site on breast cancer-specific mortality (BCSM) remains unclear. Patients and Methods We analyzed data from 305,443 women ages 20- to 79-years-old diagnosed with breast cancer between 1990 and 2009. The data were obtained from the population-based Surveillance, Epidemiology, and End Results (SEER) program of the U.S. National Cancer Institute. The survival outcomes with regard to laterality and primary tumor site were compared using univariate and multivariate (Cox proportional hazards regression model) methods. Results In the multivariate analysis, BCSM was affected by the primary tumor site (P

Girdin was identified as a novel Akt substrate that contributes to a positive feedback loop between Girdin and Akt. Although several recent studies have demonstrated that Girdin is involved in tumor metastasis, the clinical implications of Girdin in breast cancer remain unclear. To retrospectively evaluate the prognostic value of Girdin in breast cancer, we performed an immunohistochemistry screening for Girdin using tissue microarrays constructed from 250 patients who were histologically confirmed as having invasive ductal breast carcinoma at the Fudan University Shanghai Cancer Center. Our results demonstrated that the levels of Girdin in different subcellular distributions, including Girdin in the nucleus (GN) and the cytoplasm (GC) were each associated with the clinical parameters of breast cancer, including phospho-Akt (S473) [p = 0.014 for GN], phospho-Akt (T308) [p = 0.045 for GC], estrogen receptor (ER) [p = 0.012 for GN and p = 0.004 for GC], progesterone receptor (p = 0.028 for GC) and human epidermal growth factor receptor 2 status (p = 0.004 for GC). Moreover, we showed that elevated expression of GN indicated a worse disease-free survival (p = 0.032) and overall survival (p = 0.011) exclusively in the ER-positive breast cancer population. Cumulatively, our findings suggest that GN might serve as an important prognostic factor for ER-positive breast carcinoma.

BACKGROUND: Postmastectomy breast reconstruction is widely used in breast cancer patients for its aesthetic effect. Although several studies have casted suspicion upon the oncological safety of immediate breast reconstruction after mastectomy, the potential impact of different reconstruction methods on patient survival remains unclear. PATIENTS AND METHODS: We identified 35,126 female patients diagnosed with breast cancer from January 1, 1998 to December 31, 2002 in the Surveillance, Epidemiology, and End Results database. Breast cancer-specific survival (BCSS) and overall survival (OS) were compared among patients who underwent mastectomy with or without immediate breast reconstruction (autologous reconstruction or implant reconstruction) using Cox proportional hazard regression models. RESULTS: In multivariate analysis unadjusted for family income, patients undergoing immediate postmastectomy reconstruction exhibited improved BCSS [POOLED reconstruction (any types of reconstruction): hazard ratio (HR) = 0.87, 95% confidence interval (CI) 0.80-0.95, P = 0.001] and OS (pooled reconstruction: HR = 0.70, 95% CI 0.65-0.75, P

Fetoplacental endothelial cells are exposed to oxygen levels ranging from 2% to 8% in vivo. However, little is known regarding endothelial function within this range of oxygen because most laboratories use ambient air (21% O2) as a standard culture condition (SCN). We asked whether human umbilical artery endothelial cells (HUAECs) that were steadily exposed to the physiological chronic normoxia (PCN, 3% O2) for ∼20–25 days differed in their proliferative and migratory responses to FGF2 and VEGFA as well as in their global gene expression compared with those in the SCN. We observed that PCN enhanced FGF2- and VEGFA-stimulated cell proliferation and migration. In oxygen reversal experiments (i.e., when PCN cells were exposed to SCN for 24 h and vice versa), we found that preexposure to 21% O2 decreased the migratory ability, but not the proliferative ability, of the PCN-HUAECs in response to FGF2 and VEGFA. These PCN-enhanced cellular responses were associated with increased protein levels of HIF1A and NOS3, but not FGFR1, VEGFR1, and VEGFR2. Microarray analysis demonstrated that PCN up-regulated 74 genes and down-regulated 86, 14 of which were directly regulated by hypoxia-inducible factors as evaluated using in silico analysis. Gene function analysis further indicated that the PCN-regulated genes were highly related to cell proliferation and migration, consistent with the results from our functional assays. Given that PCN significantly alters cellular responses to FGF2 and VEGFA as well as transcription in HUAECs, it is likely that we may need to reexamine the current cellular and molecular mechanisms controlling fetoplacental endothelial functions, which were largely derived from endothelial models established under ambient O2.

Among chemotherapeutic agents, paclitaxel has shown great efficacy against breast cancer. Prediction of paclitaxel response may improve patient outcomes. Here we show, using exome sequencing, that in comparison with pre-treatment biopsies, two TEKT4 germline variations are enriched in post-treatment tumours. We find TEKT4 variations in ~ 10% of an independent cohort of 84 pairs of samples. Tektin4 (encoded by TEKT4) associates closely with tubulin in doublet microtubules and helps stabilize these structures. These two TEKT4 germline variations in a high cis linkage are biologically relevant, as the ectopic expression of variant TEKT4 deregulates the microtubule stability, antagonizes the paclitaxel-induced stabilizing effect of microtubules and increases paclitaxel resistance. Furthermore, TEKT4 germline variations are associated with reduced disease-free survival and overall survival compared with wild-type TEKT4 in patients undergoing paclitaxel-based chemotherapy. Taken together, we reveal a potential mechanism of resistance to paclitaxel through the acquisition of germline variations in breast cancer.

Fetoplacental endothelial cells are exposed to oxygen levels ranging from 2% to 8% in vivo. However, little is known regarding endothelial function within this range of oxygen because most laboratories use ambient air (21% O2) as a standard culture condition (SCN). We asked whether human umbilical artery endothelial cells (HUAECs) that were steadily exposed to the physiological chronic normoxia (PCN, 3% O2) for ∼20–25 days differed in their proliferative and migratory responses to FGF2 and VEGFA as well as in their global gene expression compared with those in the SCN. We observed that PCN enhanced FGF2- and VEGFA-stimulated cell proliferation and migration. In oxygen reversal experiments (i.e., when PCN cells were exposed to SCN for 24 h and vice versa), we found that preexposure to 21% O2 decreased the migratory ability, but not the proliferative ability, of the PCN-HUAECs in response to FGF2 and VEGFA. These PCN-enhanced cellular responses were associated with increased protein levels of HIF1A and NOS3, but not FGFR1, VEGFR1, and VEGFR2. Microarray analysis demonstrated that PCN up-regulated 74 genes and down-regulated 86, 14 of which were directly regulated by hypoxia-inducible factors as evaluated using in silico analysis. Gene function analysis further indicated that the PCN-regulated genes were highly related to cell proliferation and migration, consistent with the results from our functional assays. Given that PCN significantly alters cellular responses to FGF2 and VEGFA as well as transcription in HUAECs, it is likely that we may need to reexamine the current cellular and molecular mechanisms controlling fetoplacental endothelial functions, which were largely derived from endothelial models established under ambient O2.

The GATA3 gene (GATA-binding protein 3) is one of the most frequently mutated genes in breast cancer. The objective of the current study was to determine the clinicopathologic characteristics of patients with breast cancer harboring GATA3 mutations.The authors examined the somatic mutation status of GATA3 and performed survival analysis in The Cancer Genome Atlas (TCGA) cohort (n=934) and the Fudan University Shanghai Cancer Center (FUSCC) cohort (n=308). Patient characteristics, including age; menopausal status; tumor laterality; tumor size; lymph node status; tumor grade; molecular subtypes; adjuvant radiotherapy, chemotherapy, and endocrine therapy; and prognosis, together with PIK3CA (phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha) and TP53 (tumor protein p53) mutation status, were collected.GATA3 mutations were detected in 8.8% of patients (82 of 934 patients) in the TCGA cohort and 14.9% of patients (46 of 308 patients) in the FUSCC cohort. GATA3 mutations were found to be significantly associated with luminal-like breast cancer (P=.002 in the TCGA cohort and P.001 in the FUSCC cohort), and were highly mutually exclusive to PIK3CA mutations (P=.001 in the TCGA cohort and P=.003 in the FUSCC cohort) and TP53 mutations (P.001 in both cohorts). Furthermore, GATA3 mutations were correlated with improved overall survival in the entire population (P=.025 in the TCGA cohort and P = .043 in the FUSCC cohort) as well as in patients with luminal-like disease who received adjuvant endocrine therapy.GATA3 mutations mainly occur in patients with luminal-like breast cancer and have identifiable clinicopathologic and genetic characteristics, highlighting a subgroup of patients with breast cancer in whom limited therapy may be appropriate.

The aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor mediates many biological processes. Herein, we investigated if 2-(1’H-indole-3’-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE, an endogenous AhR ligand) regulated proliferation and migration of human ovarian cancer cells via AhR. We found that AhR was widely present in many histotypes of ovarian cancer tissues. ITE suppressed OVCAR-3 cell proliferation and SKOV-3 cell migration in vitro, which were blocked by AhR knockdown. ITE also suppressed OVCAR-3 cell growth in mice. These data suggest that the ITE might potentially be used for therapeutic intervention for at least a subset of human ovarian cancer.

Endothelial cells chronically reside in low-O2 environments in vivo (2%–13% O2), which are believed to be critical for cell homeostasis. To elucidate the roles of this physiological chronic normoxia in human endothelial cells, we examined transcriptomes of human umbilical vein endothelial cells (HUVECs), proliferation and migration of HUVECs in response to fibroblast growth factor 2 (FGF2) and vascular endothelial growth factor A (VEGFA), and underlying signaling mechanisms under physiological chronic normoxia. Immediately after isolation, HUVECs were cultured steadily under standard cell culture normoxia (SCN; 21% O2) or physiological chronic normoxia (PCN; 3% O2) up to 25 days. We found that PCN up-regulated 41 genes and down-regulated 21 genes, 90% of which differed from those previously reported from HUVECs cultured under SCN and exposed to acute low O2. Gene ontology analysis indicated that PCN-regulated genes were highly related to cell proliferation and migration, consistent with the results from benchtop assays that showed that PCN significantly enhanced FGF2- and VEGFA-stimulated cell proliferation and migration. Interestingly, preexposing the PCN cells to 21% O2 up to 5 days did not completely diminish PCN-enhanced cell proliferation and migration. These PCN-enhanced cell proliferations and migrations were mediated via augmented activation of MEK1/MEK2/ERK1/ERK2 and/or PI3K/AKT1. Importantly, these PCN-enhanced cellular responses were associated with an increase in activation of VEGFR2 but not FGFR1, without altering their expression. Thus, PCN programs endothelial cells to undergo dramatic changes in transcriptomes and sensitizes cellular proliferative and migratory responses to FGF2 and VEGFA. These PCN cells may offer a unique endothelial model, more closely mimicking the in vivo states.

The importance of building disaster protection is conceivable given its current situation. From the perspective of the safety ergonomics of building protection system and based on fuzzy evaluation method, this paper analyzes various factors that effect building disaster protection through Analytic Hierarchy Process (hereinafter referred to as AHP) via establishing a disaster prevention model. The study reveals that fuzzy AHP applies to security evaluation of building disaster protection, and it can provide scientific basis for design and administrative agencies to improve the current building security condition, rendering building design and operation more and more scientific.

Hemodynamics creates a constantly changing physical and chemical environment to which the arterial endothelium is exquisitely sensitive. Biomechanical stresses are intrinsic to blood flow characteristics and blood pressure and therefore are important considerations in hypertension. Near branching anatomical sites in arteries, blood flow separates from the main flow to undergo complex multi-directional characteristics for a part of each cardiac cycle (collectively referred to as disturbed flow). Atherosclerosis and aneurysmal pathology develop preferentially at disturbed flow locations, particularly when an additional cardiovascular risk factor such as hypercholesterolemia or high blood pressure are present. In the pre-disease state, a pre-inflammatory endothelial phenotype is characteristically present in such regions in contrast to nearby endothelium outside of the complex disturbed flow fields. A regulatory hierarchy of mechanisms controls endothelial gene and protein expression (and function) in response to the flow environment. Gene expression regulation, the transcription of the DNA (genomic) code to mRNA, is well described by transcription factors, enhancers and repressors, many of which are flow responsive. However, emerging studies describe the additional role of localized disturbed blood flow upon epigenetic mechanisms of endothelial responses to biomechanical stress. Epigenetics and epigenomics (global analyses of epigenetic changes across the entire genome) encompass heritable and non-heritable changes in nuclear chromatin leading to gene expression changes that cannot be attributed to changes in the primary DNA sequence. Hemodynamic stimuli have recently been shown to induce epigenetic responses including transcriptional regulation by proximal promoter DNA methylation, and regulation of gene and protein expression by histone/chromatin remodeling and by noncoding RNA-based mechanisms.Dynamic epigenomic responses to flow that result in regulation of endothelial gene expression in vivo and in vitro include: (i) regions of stable differentially methylated DNA in swine and mouse endothelial genomes that are arterial site-specific and map to atherosusceptibility, (ii) disturbed flow (but not undisturbed flow) applied to endothelial cells in vitro that induces DNA methylation through DNA methyltransferase enrichment of gene promoter regions; the resulting hypermethylation suppresses gene expression, (iii) histone acetylation / deacetylation and methylation that create histone marks that enable or suppress gene expression by controlling access of transcription factors to chromatin DNA, and (iv) short microRNAs and long noncoding RNAs that interact with highly specific binding sites of newly synthesized mRNA to promote its degradation and thus suppress transcription. Flow-mediated epigenomic responses intersect with cis and trans factor regulation to maintain endothelial function in a shear-stressed or pressure-stressed environment and may contribute to localized endothelial dysfunctions that promote localized vascular pathology.The Encyclopedia of DNA Elements (ENCODE) and the International Human Epigenome Consortia (IHEC) have uncovered thousands of putative epigenetic regulatory sites in non-coding regions of the genome. Methylation of cytosine nucleosides in DNA both at/near gene promoter regions as well as at some distance upstream is a potent epigenetic suppressor of transcription. Recently, complete methylated DNA immunoprecipitation sequencing (MeDIP-seq) of regions of arterial endothelium associated with disturbed vs undisturbed flow in vivo in pigs has identified many differentially methylated DNA profiles enriched in exons and 5'UTR sequences of annotated genes, 60 of which are linked to cardiovascular disease. Furthermore, in human arterial endothelial cells in culture we have demonstrated DNA methylation plasticity to be regulated by disturbed flow resulting in suppression (hypermethylation) or stimulation (hypomethylation) of transcription. We propose dynamic DNA methylation to be a genome-wide epigenetic mechanism of transcriptional control that regulates adaptive endothelial phenotype plasticity in response to the local environment, including biomechanical stresses associated with disturbed flow characteristics and hypertension.The epigenomic code therefore has the potential to figure prominently in disease susceptibility and pathogenesis arising from environmental influences independent of, or in concert with, mutations and single nucleotide polymorphisms (SNP) linked to many complex diseases, including hypertension and cardiovascular disease. The role of disturbed flow as a hemodynamic regulator of epigenomic DNA methylation-mediated and histone-mediated gene expression may play a prominent role in endothelial vascular function and dysfunction in hypertension, atherogenesis and aneurysm development. The cellular mechanisms that link hemodynamics and biomechanical stress to the epigenomic code remain unexplored.

Purpose To determine whether and how the patterns of breast cancer-specific mortality (BCSM) changed along with time periods. Methods We used the Surveillance, Epidemiology and End Results registry to identify 228209 female patients diagnosed with invasive breast cancer between 1990 and 2000 (cohort 1 [C1], 112981) and between 2001 and 2005 (cohort 2 [C2], 115228). BCSM was compared in two cohorts using Cox proportional hazard regression models. We analysed the relative hazard ratios (HRs) and absolute BCSM rates by flexible parametric survival modelling. Results The patterns of BCSM were similar between the two cohorts, with the peak of mortality presenting in the first 2–3 years after diagnosis, and mortality rate significantly decreased in C2 in all cases. In C2, the annual BCSM rate of all cases was 9.64 (per 1000 persons per year) in year 10 with a peak rate of 23.34 in year 2. In ER-negative and high-risk patients, marked survival improvements were achieved mostly in the first 5 years, while in ER-positive and low-risk patients, survival improvements were less but constant up to 10 years. Conclusion There has been a significant improvement of BCSM with substantially decreased mortality within 5 years. The current pattern of BCSM and its changing feature differs according to ER status. Our findings have some clinical implications both for treatment decisions and adjuvant treatment trial design.

To examine the relationship between large tumor size and breast cancer-specific mortality (BCSM), especially in a subset of patients with negative lymph nodes (LNs).We used the Surveillance, Epidemiology and End Results registry to identify 107,705 female patients diagnosed from January 1, 1990, through December 31, 2003, as having invasive breast cancer and treated with surgery and LN dissection. Relevant issues unclear in the database were studied in an additional 335 patients with locally advanced disease treated with neoadjuvant chemotherapy.In the multivariable analysis, a significant interaction was found between tumor size and LN involvement (P.001). In LN-negative diseases, the relationship between tumor size and BCSM was piecewise. Using 21- to 30-mm tumors as the reference, the hazard ratio (HR) of BCSM increased with increasing tumor size until a peak at 41 to 50 mm (HR, 1.49; P.001), after which increasing tumor size was unexpectedly related to decreasing hazard, with a nadir at 61 to 80 mm (HR, 1.06; P=.70). The 61- to 80-mm tumors exhibited a significantly lower BCSM compared with the 41- to 50-mm (P=.02) and greater than 80-mm (P=.03) subgroups. This pattern remained after stratification by estrogen receptor status but was not observed in patients with LN-positive disease. Further analysis indicated that the survival advantage of 61- to 80-mm tumors in LN-negative disease might result from its low risk of distant metastasis.A relatively larger tumor size without LN involvement may be a surrogate for biologically indolent disease of distant metastasis. Our findings, if validated in other large databases, may provide better understanding of breast cancer biology.

The Slit glycoproteins and their Roundabout (Robo) receptors regulate migration and growth of many types of cells including human cancer cells. However, little is known about the expression and roles of Slit/Robo in human ovarian cancer. Herein, we examined the expression of Slit/Robo in human normal and malignant ovarian tissues and its potential participation in regulating migration and proliferation of human ovarian cancer cells using two ovarian cancer cell lines, OVCAR-3 and SKOV-3. We demonstrated that Slit2/3 and Robo1 were immunolocalized primarily in stromal cells in human normal ovaries and in cancer cells in many histotypes of ovarian cancer tissues. Protein expression of Slit2/3 and Robo1/4 was also identified in OVCAR-3 and SKOV-3 cells. However, recombinant human Slit2 did not significantly affect SKOV-3 cell migration, and OVCAR-3 and SKOV-3 cell proliferation. Slit2 also did not induce ERK1/2 and AKT1 phosphorylation in OVCAR-3 and SKOV-3 cells. The current findings indicate that three major members (Slit2/3 and Robo1) of Slit/Robo family are widely expressed in the human normal and malignant ovarian tissues and in OVCAR-3 and SKOV-3 cells. However, Slit/Robo signaling may not play an important role in regulating human ovarian cancer cell proliferation and migration.

Histone H2A and H2B ubiquitination represents a widely used mechanism for a variety of regulatory transcriptional programs. In this review, structural and functional studies of histone H2A and histone H2B deubiquitinase (DUB), DUB including 2A-DUB, BRCA1-associated protein-1, USP3, UBP8, and USP16, and their role in developmental disease and carcinogenesis were recapitulated. Also the progress in developing small molecular inhibitors targeting DUBs and their application in colon cancer, B-cell lymphoma, and multiple myeloma were summarized. Overall, the study seek to strengthen the understanding on how these DUBs contribute to normal and malignant tissue development thus aiding in improving the design of therapeutic strategies used for diagnosis and prognosis of the disease.