β2-adrenergic receptor (β2-AR) is widely expressed on immune cells, including T cells and it has a non-canonical signaling pathway, which is β2-AR-β-arrestin 2 (β-Arr2)-extracellular signal-regulated kinase 1/2 (ERK1/2). Our previous studies have shown that the β2-AR agonist terbutaline (Terb) can activate β2-AR and significantly inhibit helper T (Th) 17 cell function in collagen-induced arthritis (CIA). However, the effects of β2-AR on regulatory T (Treg) cells in CIA have not been consistently determined. The aim of our research was to explore whether β2-AR-β-Arr2-ERK1/2 signaling in Treg cells regulates inflammatory responses and signs in CIA. DBA1/J mice were used to prepare CIA model by intradermal injection of collagen type II (CII). Inducible Treg (iTreg) cells were induced from CD4+ T cells that were isolated from the spleens of normal or CIA mice and treated with Terb, β-Arr2 gene silence or overexpression or the ERK1/2 inhibitor U0126 in vitro. β2-AR and β-Arr2 expression, pERK1/2 level, interleukin (IL)-10 and transforming growth factor (TGF)-β production and the suppression of effector T (Teff) cell proliferation mediated by nature Treg (nTreg) cells were determined. β-Arr2-overexpressed Treg cells were intravenously injected into the tail base of CIA mice. Arthritic symptoms were assessed by clinical arthritis scores. Frequencies of Th17 and Treg cells, cytokine production and osteoclast and osteoblast-specific gene expression were estimated. The increased β-Arr2 expression and pERK1/2 level induced by Terb in CIA iTregs were reduced by β-Arr2 gene silence. The increased TGF-β and IL-10 production in iTreg cells by Terb and the suppression of Teff cell proliferation mediated by Terb-treated nTregs from CIA mice were downregulated by β-Arr2 gene silence or U0126 and further enhanced by β-Arr2 gene overexpression. Adoptive transfer of β-Arr2-overexpressed Treg cells into CIA mice reduced limb inflammation, osteoclast-specific gene expression in joints and Th17 cell cytokine production in joints and serum but increased osteoblast-specific gene expression in joints and Treg cell cytokine production in joints and serum. β2-AR/β-Arr2/ERK signaling in Treg cells contributes to alleviation of inflammatory responses and symptoms in CIA.

MDSCs in breast cancer tissues

MDSCs produce NO and IL-6 and induce tumor STAT3 phosphorylation

Correlations between CD33, ALDH1A1 and IL6 transcripts in patients with breast cancer

Supplementary tables: Supplementary Table 1. Patient characteristics (All 3 cohorts) Supplementary Table 2. shRNA Sequences and Real-time PCR primers Supplementary Table 3. Multivariate analysis of overall survival (All 3 cohorts) Supplementary Table 4. Multivariate analysis of overall survival (Cohort 1, HER2+ patients) Supplementary Table 5. Multivariate analysis of overall survival (Cohorts 2 and 3, random patients)

MDSC phenotype and suppressor activity

Relationship between MDSCs and breast cancer stemness

Supplementary Figure 1 MDSC phenotype and suppressor activity Supplementary Figure 2 MDSCs in breast cancer tissues Supplementary Figure 3 Relationship between MDSCs and breast cancer stemness Supplementary Figure 4 MDSCs produce NO and IL-6 and induce tumor STAT3 phosphorylation Supplementary Figure 5 Correlations between CD33, ALDH1A1 and IL6 transcripts in patients with breast cancer

Esophageal squamous cell carcinoma (ESCC) is a common malignant tumour type of the digestive system. Long non-coding RNA (lncRNA) c-Myc upregulated (MYU), also known as VPS9 domain-containing 1 antisense 1, was recently discovered. However, the expression of lncRNA MYU in ESCC and its role in tumour progression have remained elusive. In the present study, the expression of lncRNA MYU, Ki-67 and the epithelial-mesenchymal transition-related proteins E-cadherin and Vimentin in ESCC tissues was detected by reverse transcription-quantitative PCR. The expression of Ki-67, E-cadherin and Vimentin in ESCC tissues was also detected by immunohistochemistry. A small interfering RNA plasmid was employed to establish a TE-2 cell line with knockdown on lncRNA MYU. The results indicated that the expression of lncRNA MYU was higher in ESCC tissues than in normal adjacent tissues and that upregulation of lncRNA MYU was a potential biomarker for poor prognosis. The results also suggested that the expression levels of lncRNA MYU were correlated with the histological grade, lymph node metastasis and TNM stage (P<0.05). Silencing of lncRNA MYU expression inhibited the proliferation, migration and invasion, while the expression of lncRNA MYU increased as cell proliferation increased. In addition, the mRNA expression of Vimentin and Ki-67 was decreased in TE-2 cells after lncRNA MYU was knocked down, while E-cadherin mRNA expression was elevated. In conclusion, the present results indicated that lncRNA MYU may regulate the proliferation, migration and invasion of ESCC cells, and may serve as a prognostic biomarker for ESCC.

Cisplatin (CDDP) has been extensively used for gastric cancer (GC) treatment but limited by drug resistance and severe toxicity.The chemo-sensitizers that enhance its efficiency and overcome its limitation are urgently needed.Oxymatrine (OMT), a primary active ingredient from the dry roots of Sophora favescens, has shown powerful anti-cancer property with little side-effect.In this study, we explored the chemo-sensitization of OMT to potentiate the anti-tumor effect of CDDP.GC cell lines were dealt with OMT and/or CDDP and then subjected to different experimental methods.We found that OMT could significantly potentiate the CDDP-caused BGC-823 and SGC7901 cells viability loss, and OMT acts synergistically with CDDP.The combinative treatment could arrest cell cycle in G0/G1 phase by increasing p21, p27 and decreasing cyclin D1, and induced apoptosis by ROS generation and AKT/ERK inactivation.Inhibition of ROS respectively reversed the cell death induced by OMT and/or CDDP, suggesting the pivotal roles of ROS in the process.Moreover, OMT enhanced the antitumor effects of CDDP in nude mice bearing BGC823 tumor xenografts in vivo.Taken together, this study highlights that the co-treatment with OMT and CDDP exerted synergistic antitumor effects in GC cells, and that these effects may be mediated by ROS generation and inactivation of the AKT/ERK pathways.

Herein, the Attapulgite nanoparticles (ATP NPs) coated silk fabric was prepared by impregnation method using hyperbranched polymer as addition agent. The ATP NPs and prepared silk fabrics were characterized by means of scanning electron microscope (SEM), ultraviolet-visible (UV–Vis) spectroscopy, Fourier transform infrared spectrophotometer (FTIR), X-ray diffraction (XRD), Energy Dispersive Spectrometer (EDS). The results of SEM, EDS, FTIR and XRD confirmed that ATP NPs were successfully coated on the surface of silk fabric. Not only did the treated silk fabrics possess excellent antibacterial property and antibacterial resistance, but also exhibited outstanding anti-ultraviolet performance, which can meet the requirements of multifunctional products.

BACKGROUND Norepinephrine (NE), a neurotransmitter released from the sympathetic nerves, has been shown to be involved in rheumatoid arthritis (RA). However, its role in the sympathetic nervous system in RA is divergent. Herein, we demonstrate that the sympathetic neurotransmitter NE exerts an anti-inflammatory effect in collagen-induced arthritis (CIA), a mouse model of RA, by inhibiting Th17 cell differentiation and function via β2-adrenergic receptor (β2-AR) signaling. MATERIAL AND METHODS CIA was prepared by intradermal injection of collagen type II in the tail base of DBA1/J mice. On the 41st day post-immunization, the mice were used as CIA models. CD4+ T cells from the spleen were purified using magnetic cell sorting and activated with anti-CD3 anti-CD28 antibodies. Th17 cells were polarized from the CD4+ T cells using various antibodies and cytokines. RESULTS Co-expression of CD4 and β2-AR was observed in spleens of both intact and CIA mice. The β2-AR expression in the ankle and spleen was downregulated in CIA mice. CIA induced increases in production of interleukin (IL)-17 and IL-22, CD25-IL-17+ cell percentage, and ROR-γt expression in CD4+ T cells. Importantly, NE reduced the CIA-induced CD4+ T cell shift towards Th17 phenotype, and the β2-AR antagonist ICI118551 blocked the NE effect. Moreover, the β2-AR agonist terbutaline (Terb) inhibited CIA-induced CD4+ T cell proliferation and shift towards Th17 phenotype, and the protein kinase A (PKA) inhibitor H-89 abolished the agonist effect. Terb also reduced CIA-induced Th17 enhancement, and H-89 impaired the Terb effect. CONCLUSIONS NE inhibits Th17 cell differentiation and function in CIA condition by activation of β2-AR/PKA signaling.

Tyrosine hydroxylase (TH), a rate-limiting enzyme for the synthesis of catecholamines, is expressed in T lymphocytes. However, the role of T cell-expressed TH in rheumatoid arthritis (RA) is less clear. Herein, we aimed to show the contribution of TH expression by CD4+ T cells to alleviation of helper T (Th)17/regulatory T (Treg) imbalance in collagen-induced arthritis (CIA), a mouse model of RA. CIA was prepared by intradermal injection of collagen type II (CII) at tail base of DBA1/J mice. Expression of TH in the spleen and the ankle joints was measured by real-time polymerase chain reaction and Western blot analysis. Percentages of TH-expressing Th17 and Treg cells in splenic CD4+ T cells were determined by flow cytometry. Overexpression and knockdown of TH gene in CD4+ T cells were taken to evaluate effects of TH on Th17 and Treg cells in CIA. TH expression was upregulated in both the inflamed tissues (spleen and ankle joints) and the CD4+ T cells of CIA mice. In splenic CD4+ T cells, the cells expressing TH were increased during CIA. These cells that expressed more TH in CIA were mainly Th17 cells rather than Treg cells. TH gene overexpression in CD4+ T cells from CIA mice reduced Th17 cell percentage as well as Th17-related transcription factor and cytokine expression and secretion, whereas TH gene knockdown enhanced the Th17 cell activity. In contrast, TH gene overexpression increased Treg-related cytokine expression and secretion in CD4+ T cells of CIA mice, while TH gene knockdown decreased the Treg cell changes. Collectively, these findings show that CIA induces TH expression in CD4+ T cells, particularly in Th17 cells, and suggest that the increased TH expression during CIA represents an anti-inflammatory mechanism.

Foxp3(+)CD4(+) regulatory T (Treg) cells are thought to express negligible levels of effector cytokines, and inhibit immune responses and inflammation. Here, we have identified a population of IL-8(+)Foxp3(+)CD4(+) T cells in human peripheral blood, which is selectively increased in the microenvironments of ulcerative colitis and colon carcinoma. Phenotypically, this population is minimally overlapping with IL17(+)Foxp3(+)CD4(+) T cells, and is different from IL-8(-)Foxp3(+)CD4(+) T cells in the same microenvironment. 40-60% of IL-8(+)Foxp3(+)CD4(+) T cells exhibit naive phenotype and express CD127, whereas IL8(-)Foxp3(+)CD4(+) cells are basically memory T cells and express minimal CD127. The levels of CXCR5 expression are higher in IL-8(+)Foxp3(+) cells than in IL-8(-)Foxp3(+) cells. IL-2 and TGF beta induce IL-8(+)Foxp3(+) T cells. Exogenous Foxp3 expression promotes IL-8(+)Foxp3(+) T cells and inhibits effector cytokine IFN gamma and IL-2 expression. Furthermore, Foxp3 binds to IL-8 proximal promoter and increases its activity. Functionally, IL-8(+)Foxp3(+) T cells inhibit T cell proliferation and effector cytokine production, but stimulate inflammatory cytokine production in the colon tissues, and promote neutrophil trafficking through IL-8. Thus, IL-8(+)Foxp3(+) cells may be an "inflammatory" Treg subset, and possess inflammatory and immunosuppressive dual biological activities. Given their dual roles and localization, these cells may be in a unique position to support tumor initiation and development in human chronic inflammatory environment.

Abstract Myeloid-derived suppressor cells (MDSC) contribute to immune suppression in cancer, but the mechanisms through which they drive metastatic progression are not fully understood. In this study, we show how MDSC convey stem-like qualities to breast cancer cells that coordinately help enable immune suppression and escape. We found that MDSC promoted tumor formation by enhancing breast cancer cell stem-like properties as well as by suppressing T-cell activation. Mechanistic investigations indicated that these effects relied upon cross-talk between the STAT3 and NOTCH pathways in cancer cells, with MDSC inducing IL6-dependent phosphorylation of STAT3 and activating NOTCH through nitric oxide leading to prolonged STAT3 activation. In clinical specimens of breast cancer, the presence of MDSC correlated with the presence of cancer stem-like cells (CSC) and independently predicted poor survival outcomes. Collectively, our work revealed an immune-associated mechanism that extrinsically confers cancer cell stemness properties and affects patient outcome. We suggest that targeting STAT3-NOTCH cross-talk between MDSC and CSC could offer a unique locus to improve cancer treatment, by coordinately targeting a coupled mechanism that enables cancer stemness and immune escape. Cancer Res; 76(11); 3156–65. ©2016 AACR.

Recently, many studies have been published to evaluate the correlation between the cytochrome P450 1A1 (CYP1A1) T3801C polymorphism and cervical neoplasia risk. However, the results remain inconclusive. To clarify this possible association, we conducted a systematic review and meta-analysis of published studies. Data were collected from the following electronic databases: PubMed, Embase, Ovid, ISI Web of Knowledge, Google Scholar, and Chinese Biomedical Database databases. The pooled odds ratio (OR) and its 95 % confidence interval (95 % CI) were used to assess the strength of this association. The pooled ORs were performed for the allele model (C vs. T), the homozygote model (CC vs. TT), the dominant model (CC/CT vs. TT), and the recessive model (CC vs. TT/CT), respectively. Finally, a total of 12 independent studies including a total of 3,724 subjects (1,912 cases/1,812 controls) were eligible for meta-analysis. Overall, there was a significant association between the CYP1A1 T3801C polymorphism and cervical neoplasia susceptibility (C vs. T, OR 1.32, 95 % CI 1.04–1.68, P = 0.02; CC vs. TT, OR 1.99, 95 % CI 1.19–3.35, P = 0.01; CC/CT vs. TT, OR 1.36, 95 % CI 1.02–1.81, P = 0.02; CC vs. TT/CT, OR 1.57, 95 % CI 1.23–2.02, P < 0.01). Meta-analysis of the ten studies on cervical cancer suggested a significant association between the CYP1A1 T3801C polymorphism and cervical cancer risk (C vs. T, OR 1.38, 95 % CI 1.05–1.82, P = 0.02; CC vs. TT, OR 2.06, 95 % CI 1.15–3.70, P = 0.02; CC/CT vs. TT, OR 1.45, 95 % CI 1.03–2.02, P = 0.03; CC vs. TT/CT, OR 1.56, 95 % CI 1.20–2.03, P < 0.01). In the stratified analysis by ethnicity, significant associations were also detected in some genetic models. This meta-analysis demonstrates a significant association between the CYP1A1 T3801C polymorphism and cervical neoplasia susceptibility.

Tumor-associated macrophages (TAMs), most of which exhibit M2 phenotype, function as immunosuppressive cells in tumor microenvironment (TME). Polarization of TAMs from a pro-immune (M1 like) phenotype to an immune-suppressive (M2-like) phenotype is one of the hallmarks of malignancy, but their molecular basis is still remains unknown. It has been reported that microRNAs are involved in monocyte-macrophage differentiation. In this study, we found that miR-19a-3p, broadly conserved in vertebrate, could reverse the M2 phenotype of RAW macrophage cells. When mouse breast tumor cells such as 4T1, 4TO7 and EMT6 were co-cultured with RAW macrophage cells which over express miR-19a-3p, the invasion capacity was suppressed. Meanwhile, when the conditional medium of RAW cells which were transfected with miR-19a-3p mimic was added into culturing medium of tumor cells, the migration capacity of 4T1 and EMT6 breast cancer cells was inhibited. Moreover, when miR-19a-3p was injected intratumor, consistent with the in vitro experiments, we found that the M2 phenotype of TAMs was suppressed significantly. Although 4T1 xengraft breast tumor growth was not affected by miR-19a-3p, lung metastasis of tumor cells was significantly suppressed. Taken together, our findings indicate that miR-19a-3p is down-regulated in M2 phenotype RAW macrophage and TAMs in TME. The low expression of miR-19a-3p plays an important role in inducing M2 macrophage polarization and promoting migration and invasion capacity and metastasis. Citation Format: Jian Yang, Na Li, Zhuhong Zhang, Qin Si, Chong Chen, Yan Liu, Ralph A. Reisfeld, Peiqing Sun, Dwayne Stupack, Rong Xiang, Yunping Luo. miR-19a-3p inhibits breast carcinoma metastasis via reversing M2 phenotype of TAMs. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2828. doi:10.1158/1538-7445.AM2013-2828 Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.

OBJECTIVE To investigate the negative regulation of microRNA-1 (miR-1) on L-type calcium channel beta2 subunit (Cavbeta 2) during cardiomyocyte hypertrophy and its mechanism. METHODS Cardiomyocyte hypertrophy was induced by isoproterenol (ISO). The cell surface area was measured by image analysis system (HJ2000). The targets of miR-1 were predicted by online database microCosm. The 3' untranslated region sequence of Cavbeta 2 was cloned into luciferase reporter vector and then transiently transfected into HEK293 cells. The luciferase activities of samples were measured to verify the expression of luciferase reporter vector. The expression of atrial natriuretic peptide (ANP), beta-myosin heavy chain (beta-MHC), miR-1 and the Cavbeta 2 mRNA were detected by qRT-PCR. The protein expression of Cavbeta 2 was detected by Western blot. The level of miR-1 was up-regulated by miR-1 mimic transfection and the expression level of Cavbeta 2 was down-regulated by RNAi, then effects of which on cardiomyocyte hypertrophy were investigated. RESULTS (1) The expression of miR-1 was significantly reduced in cardiomyocyte hypertrophy. Upregulating the miR-1 level could suppress the increase of cell surface area, the expression of ANP and beta-MHC mRNA (P < 0.05). (2) Cavbeta 2 was the one of potential targets of miR-1 by prediction using online database microCosm. The luciferase activities of HEK293 cells with the plasmid containing miR-1 and wide type Cavbeta 3' UTR sequence was significantly decreased when compared with that of control group (P < 0.01). Up-regulation of the miR-1 level could suppress the protein expression of Cavbeta 2. (3) The expression of Cavbeta 2 was significantly increased in cardiomyocyte hypertrophy induced by ISO. Downregulation of Cavbeta by RNAi could markedly inhibit the increase of cell surface area, the expression of ANP and beta-MHC mRNA. CONCLUSION Cavbeta2 is one of potential targets of miR-1 by bioinformatics prediction. The experiment data confirms that Cavbeta2 is truly the target of miR-1. MiR-1 can negatively regulate the expression of Cavbeta 2, resulting in the decrease of intracellular Ca2+ content and the attenuation of cardiomyocyte hypertrophy.

OBJECTIVE:To investigate the expressions of Fas, NF-kappaB, and vascular endothelial growth factor-C (VEGF-C) in infiltrating ductal carcinoma of the breast and provide scientific basis for early diagnosis and prognosis of breast cancer. MATERIALS AND METHODS:The immunohistochemical technique (SP method) was used to detect expression of Fas, NF-kappaB, and VEGF-C in 137 cases of breast-infiltrating ductal carcinoma, 17 cases of intraductal carcinoma of the breast, and 20 cases of normal breast tissues, and analyze its relationship with clinicopathologic factors of breast cancer and patients' survival rate, as well as the correlation among their expression, clinicopathologic factors, and survival rate. RESULTS:Fas expression was less commonly detected in infiltrating ductal carcinoma than in intraductal carcinoma and normal tissue. In contrast, both NF-kappaB and VEGF-C were more commonly detected in infiltrating ductal carcinoma than in intraductal carcinoma and normal tissue. Fas expression was correlated with tumor size, histological grade, and clinicopathological stage; NF-kappaB expression was correlated with tumor size, histological grade, lymph node metastasis; VEGF-C expression was correlated with lymph node metastasis and clinical and pathological stages of breast cancer (p < or = 0.05). Spearman rank correlation analysis revealed a negative correlation between Fas expression and both NF-kappaB and VEGF-C expression in infiltrating breast cancer (p < 0.05) Additionally, Kaplan-Meier survival analysis demonstrated that five-year survival was higher for patients with Fas-positive samples but lower for those with VEGF-C-positive samples. CONCLUSIONS:The present results demonstrate that Fas and NF-B play a role in the initiation and development of breast cancer, while VEGF-C appears to promote lymph node metastasis. Thus, these proteins may serve as useful diagnostic and prognostic markers of invasive breast cancer.