BackgroundToll-like receptors (TLR) are key innate immunity receptors participating in an immune response. Growing evidence suggests that mutations of TLR2/TLR9 gene are associated with the progress of cancers. The present study aimed to investigate the temporal relationship of single nucleotide polymorphisms (SNP) of TLR2/TLR9 and the risk of hepatocellular carcinoma (HCC).MethodsIn this single center-based case-control study, SNaPshot method was used to genotype sequence variants of TLR2 and TLR9 in 211 patients with HCC and 232 subjects as controls.ResultsTwo synonymous SNPs in the exon of TLR2 were closely associated with risk of HCC. Compared with those carrying wild-type homozygous genotypes (T/T), risk of HCC decreased significantly in individuals carrying the heterozygous genotypes (C/T) of the rs3804099 (adjusted odds ratio (OR), 0.493, 95% CI 0.331 - 0.736, P < 0.01) and rs3804100 (adjusted OR, 0.509, 95% CI 0.342 - 0.759, P < 0.01). There was no significant association found in two TLR9 SNPs concerning the risk of HCC. The haplotype TT for TLR2 was associated significantly with the decreased risk of HCC (OR 0.524, 95% CI 0.394 - 0.697, P = 0.000). Inversely, the risk of HCC increased significantly in patients with the haplotype CC (OR 2.743, 95% CI 1.915 - 3.930, P = 0.000).ConclusionsThese results suggested that TLR2 rs3804099 C/T and rs3804100 C/T polymorphisms were closely associated with HCC. In addition, the haplotypes composed of these two TLR2 synonymous SNPs have stronger effects on the susceptibility of HCC.
beta-Elimination of the phosphate group on phosphoserine and phosphothreonine residues and addition of an alkyldithiol is a useful tool for analysis of the phosphorylation states of proteins and peptides. We have explored the influence of several conditions on the efficiency of this PO(4)(3-) elimination reaction upon addition of propanedithiol. In addition to the described influence of different bases, the solvent composition was also found to have a major effect on the yield of the reaction. In particular, an increase in the percentage of DMSO enhances the conversion rate, whereas a higher amount of protic polar solvents, such as water or isopropanol, induces the opposite effect. We have also developed a protocol for enrichment of the modified peptides, which is based on solid-phase covalent capture/release with a dithiopyridino-resin. The procedure for beta-elimination and isolation of phosphorylated peptides by solid-phase capture/release was developed with commercially available alpha-casein. Enriched peptide fragments were characterized by MALDI-TOF mass spectrometric analysis before and after alkylation with iodoacetamide, which allowed rapid confirmation of the purposely introduced thiol moiety. Sensitivity studies, carried out in order to determine the detection limit, demonstrated that samples could be detected even in the low picomolar range by mass spectrometry. The developed solid-phase enrichment procedure based on reversible covalent binding of the modified peptides is more effective and significantly simpler than methods based on the interaction between biotin and avidin, which require additional steps such as tagging the modified peptides and work-up of the samples prior to the affinity capture step.
The aim of the current study was to investigate the regulatory effect of miR-146a on the toll-like receptor 4 (TLR-4)/NF-κB pathway and therefore inflammation in septic cardiomyopathy. A total of 60 healthy male Sprague Dawley rats were equally divided into a control, LPS, miR-146a agonist and miR-146a inhibitor group. Blood samples were collected from rats 24 h after intraperitoneal lipopolysaccharide injection and myocardial tissues were subsequently collected. After hematoxylin and eosin staining of rat myocardial tissues, the degree of inflammatory cell infiltration and myocardial damage was observed. The content of certain myocardial injury markers were also observed, including cardiac troponin I (cTnI), B-type natriuretic peptide (BNP), creatine kinase myocardial bound (CK-MB) and myoglobin (Mb). Western blot analysis was performed to detect the expression of NF-κB/TLR-4, tumor necrosis factor (TNF-α) and intercellular adhesion molecule-1 (ICAM-1) in myocardial tissues. Reverse transcription-quantitative (RT-q) PCR was used to detect the expression of miR-146a, TNF-α, interleukin (IL)-1α and IL-1β mRNA in myocardial tissues. In the LPS group, myocardial interstitial tissue edema occurred, with enlarged and loosely arranged cardiomyocytes. Compared with the sepsis model group, myocardial interstitial tissue edema was relieved in the miR-146a agonist group, but was aggravated in the miR-146a inhibition group. The serum levels of cTnI, BNP, CK-MB, Mb, NF-κB, TLR-4, TNF-α and ICAM-1 in the sepsis model group were higher than those in the control group. In the miR-146a agonist group, levels of myocardial injury markers were lower than those in the sepsis model group, but were higher in the miR-146a inhibition group. The results of RT-qPCR demonstrated that the expression of miR-146a, TNF-α, IL-1α and IL-1β in the sepsis model group were upregulated compared with the control group. In addition, miR-146a expression in the miR-146a agonist group and the miR-146a inhibition group was increased, but TNF-α, IL-1α and IL-1β mRNA was downregulated. miR-146a may regulate the TLR-4/NF-κB signaling pathway via negative feedback mechanisms, leading to the improvement of the inflammatory response and cardiac dysfunction in sepsis-induced cardiomyopathy.
The present study aimed to evaluate the expression of hypoxia-inducible factor-1α (HIF-1α) and MDR1/P-glycoprotein (P-gp) in human laryngeal squamous cell carcinoma (LSCC) tissues, and also to investigate the regulation of MDR1 gene expression by HIF-1α in Hep-2 cells under hypoxic conditions. The expression of HIF-1α and MDR1/P-gp in human LSCC tissues was examined using immunohistochemistry. The HIF-1α and MDR1 gene expression in the Hep-2 cells was detected using real-time quantitative reverse transcription (QRT)-PCR and western blot analysis under normoxic and hypoxic conditions. In hypoxia, HIF-1α expression was inhibited by RNA interference. HIF-1α and MDR1/P-gp expression was high in the LSCC tissues and was associated with the clinical stage and lymph node metastasis (P<0.05). HIF-1α expression was positively correlated with MDR1/P-gp expression (P<0.01). In the Hep-2 cells, HIF-1α and MDR1/P-gp expression significantly increased in response to hypoxia. The inhibition of HIF-1α expression synergistically downregulated the expression of the MDR1 gene in hypoxic Hep-2 cells. HIF-1α expression is positively correlated with MDR1/P-gp expression in LSCC, and the two proteins may be able to serve as potential biomarkers for predicting the malignant progression and metastasis of LSCC. HIF-1α may be critical for the upregulation of MDR1 gene expression induced by hypoxia in Hep-2 cells.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.