Low Expression of Toll-like Receptor 4 Is Associated With Poor Prognosis in Bladder Cancer
Yoshito KusuharaKei DaizumotoKAICHI KAWAIKento HirayamaMinoru KowadaTerumichi ShintaniYayoi FukuharaTsogt-Ochir DondooKeisuke OzakiMegumi TsudaTomoya FukawaHiroyoshi NakatsujiYoshimi BandoHisanori UeharaTomoharu FukumoriMasayuki TakahashiHiro‐omi Kanayama
9
Citation
26
Reference
10
Related Paper
Citation Trend
Abstract:
Background/Aim: The aim of this study was to elucidate the relationship between the progression of bladder cancer (BCa) and TLR4 expression. Materials and Methods: The relationship between TLR4 expression and prognosis of BCa patients was analyzed using a publicly available database and immunohistochemical staining of clinical samples. The effect of TLR4 knockdown was also examined on the invasive capabilities of BCa cells. Finally, to investigate the biological function of TLR4, the gene expression profile of TLR4-depleted BCa cells was analyzed by microarray analysis. Results: Expression of TLR4 was inversely associated with prognosis of patients with invasive BCa, and depletion of TLR4 significantly enhanced the invasive capability of BCa cells. Gene expression profiling revealed that depletion of TLR4 led to high expression of epithelial differentiation genes. Furthermore, expression of TLR4 was found to be extremely low in areas of squamous differentiation. Conclusion: Low TLR4 expression was correlated with tumor progression.Keywords:
Tumor progression
Objective:To investigate the influences of Aβ25-35 on the TLR3 and TLR4 mRNA. Method: NG108-15 cell line was stimulated with Aβ25-35,the supernatant was collected 24 hours later and the inflammatory factors TNF-α and IFN-β were quantitatively measured,the changes in the mRNA expression of TLR3 and TLR4 were detected by real-time PCR. Result:Aβ induced the over-secreting of TNF-α、IFN-β and over-expression of TLR3 and TLR4 mRNA (P 0. 01). Conclusions:Aβ induced the over-secreting of inflammatory factors and over-expression of TLR3 and TLR4,indicating the action of Aβ in causing AD disease might be affiliated with the immune inflammation and the TLRs signaling pathway.
TLR3
Cite
Citations (0)
Cite
Citations (8)
Toll-like receptors (TLRs) are the best characterized Pathogen Recognition Receptors (PRRs) and are directly responsible for initiating an appropriate defense against bacterial and viral infection. Among all the TLRs known, only TLR4 is able to activate both MyD88-dependent induction of genes encoding inflammatory molecules and TRIF-dependent production of type I interferon. Therefore, in this study we report the binding of TLR4 by Lipopolysaccharide (LPS) which is the component on the cell-wall of gram-negative bacteria. Binding of LPS is a prerequisite for the activation of Toll-like receptor 4 (TLR4) by LPS which increases the expression of critical proinflammatory cytokines that organize potent immune responses. The binding of LPS to TLR4 was studied using IC21 mice macrophage cell line and fluorescently labeled LPS molecule, called FITC-LPS by flow cytometry. The series of cell staining experiments were performed, which included binding of FITC-LPS to TLR4 at different temperatures as temperature influences cellular trafficking of TLR4. Since, trafficking or internalization of LPS depends on its aggregation behavior; the molecular state of LPS under experimental conditions is detected using SDS-PAGE. Trypan blue was used to identify surface bond versus internalized FITC-LPS.
Internalization
Proinflammatory cytokine
TRIF
Cite
Citations (0)
Abstract Toll-like receptors (TLRs) recognize pathogen-associated molecular patterns, which are non-self macromolecular components of pathogens that allow the innate-immune system to recognize infection. TLRs are expressed on macrophages and dendritic cells (DC). TLR stimulation or CD40 agonists can induce inflammatory cytokine secretion from macrophages and DC, and promote DC maturation. The regulation of TLR expression by inflammation has begun to be explored. Our studies have focused on the regulation of TLR4 surface expression on DC. TLR4, along with the adaptor molecule MD2, is involved in the recognition of lipopolysaccharide (LPS). CD40 stimulation via cross-linked anti-CD40 monoclonal antibody (mAb) up-regulates TLR4-MD2 surface expression on a DC cell line (DC2.4) and on ex vivo-cultured splenic DC. LPS treatment down-regulated surface TLR4-MD2 on DC2.4 cells, but if combined with anti-CD40 mAb, increased TLR4-MD2 expression was observed. The increased TLR4-MD2 surface expression by any treatment did not correlate with TLR4 mRNA levels. The functional consequence of increased TLR4-MD2 expression following LPS and anti-CD40 treatment was examined. Although CD40 prestimulation did slightly enhance interleukin-12p70 secretion after LPS restimulation, simultaneous anti-CD40 mAb and LPS treatment, which up-regulates TLR4-MD2 complex, does not restore DC responsiveness to subsequent LPS.
Ex vivo
Cite
Citations (16)
This study aimed to use gene chips to investigate differential gene expression profiles in the occurrence and development of acute myocardial infarction (AMI). The study included 12 AMI patients and 12 healthy individuals. Total mRNA of peripheral bloodwas extracted and reversed-transcribed to cDNA for microarray analysis. After establishing two pools with three subjects each (3 AMI patients and 3 healthy individuals), the remaining samples were used for RT-qPCR to confirm the microarray data. From the microarray results, seven genes were randomly selected for RT-qPCR. RT-qPCR results were analyzed by the 2-ΔΔCt method. Microarray analysis showed that 228 genes were up- regulated and 271 were down-regulated (p ≤ 0.05, |logFC| > 1). Gene ontology showed that these genes belong to 128 cellular components, 521 biological processes, and 151 molecular functions. KEGG pathway analysis showed that these genes are involved in 107 gene pathways. RT-qPCR results for the seven genes showed expression levels consistent with those obtained by microarray. Thus, microarray data could be used to select the pathogenic genes for AMI. Investigating the abnormal expression of these differentially expressed genes might suggest efficient strategies for the prevention, diagnosis, and treatment of AMI.
Microarray databases
KEGG
Gene chip analysis
Cite
Citations (18)
Gene chip analysis
Microarray databases
Cite
Citations (8)
Proinflammatory cytokine
Cite
Citations (34)
Toll-like receptor 4 (TLR4) is a mammalian homologue of Drosophila Toll, a leucine-rich repeat molecule that can trigger innate responses against pathogens. The TLR4 gene has recently been shown to be mutated in C3H/HeJ and C57BL/10ScCr mice, both of which are low responders to lipopolysaccharide (LPS). TLR4 may be a long-sought receptor for LPS. However, transfection of TLR4 does not confer LPS responsiveness on a recipient cell line, suggesting a requirement for an additional molecule. Here, we report that a novel molecule, MD-2, is requisite for LPS signaling of TLR4. MD-2 is physically associated with TLR4 on the cell surface and confers responsiveness to LPS. MD-2 is thus a link between TLR4 and LPS signaling. Identification of this new receptor complex has potential implications for understanding host defense, as well as pathophysiologic, mechanisms.
Cite
Citations (2,197)
Background: LPS sensitivity in the liver significantly worsens the outcome of gram negative sepsis. MD-2 facilitates the recognition of LPS by Toll-like receptor (TLR4). In this study we investigated the changes in TLR4 and MD-2 levels in LPS sensitive livers.
Cite
Citations (0)
Abstract The human homologue of Drosophila Toll (hToll), also called Toll-like receptor 4 (TLR4), is a recently cloned receptor of the IL-1/Toll receptor family. Interestingly, the TLR4 gene has been localized to the same region to which the Lps locus (endotoxin unresponsive gene locus) is mapped. To examine the role of TLR4 in LPS responsiveness, we have generated mice lacking TLR4. Macrophages and B cells from TLR4-deficient mice did not respond to LPS. All these manifestations were quite similar to those of LPS-hyporesponsive C3H/HeJ mice. Furthermore, C3H/HeJ mice have, in the cytoplasmic portion of TLR4, a single point mutation of the amino acid that is highly conserved among the IL-1/Toll receptor family. Overexpression of wild-type TLR4 but not the mutant TLR4 from C3H/HeJ mice activated NF-κB. Taken together, the present study demonstrates that TLR4 is the gene product that regulates LPS response.
Cite
Citations (3,373)