ABSTRACT
Crytotanshinone (CTN), one of the main constituents of Salvia miltiorrhiza, has been known to exhibit antioxdative, anti-inflammatory and other important therapeutic activities. The aim of this study was to evaluate the effect of CTN on prostaglandin E2 and COX-2 production in LPS-stimulated human intestinal cells (Caco-2â¯cells). Caco-2â¯cells were stimulated with LPS in the presence or absence of CTN. The production of prostaglandin E2 (PGE2) was detected by ELISA. The expression of COX-2 was detected by qRT-PCR and Western blot. The extent of phosphorylation of IκB-α, NF-κB p65 and the expression of TLR4 were detected by western blot. The results showed that CTN dose-dependently inhibited the expression of COX-2 both in mRNA and protein levels, resulting in a decreased production of PGE2. We also found that CTN suppressed LPS-induced NF-κB activation and IκBα degradation. Furthermore, CTN inhibited the expression of TLR4 up-regulated by LPS. These results suggest that CTN exerts an anti-inflammatory property by inhibiting TLR4/NF-κB signaling pathway and the release of pro-inflammatory mediators. These findings suggest that CTN may be a therapeutic agent against intestinal inflammatory diseases.
Subject(s)
Anti-Inflammatory Agents/metabolism , Cyclooxygenase 2/metabolism , Dinoprostone/antagonists & inhibitors , Epithelial Cells/drug effects , Lipopolysaccharides/toxicity , NF-kappa B/antagonists & inhibitors , Phenanthrenes/metabolism , Toll-Like Receptor 4/antagonists & inhibitors , Anti-Inflammatory Agents/isolation & purification , Caco-2 Cells , Cyclooxygenase 2/biosynthesis , Dinoprostone/biosynthesis , Humans , Phenanthrenes/isolation & purification , Salvia miltiorrhiza/chemistry , Signal TransductionABSTRACT
Rab11-family interacting proteins (Rab11FIPs) are associated with the progression of various tumors; however, their expression and clinical significance in colorectal cancer (CRC) remains largely undetermined. In this study, the clinical implications, functions and underlying mechanisms of Rab11FIP4 in CRC were investigated. Immunohistochemical analysis revealed that expression of Rab11FIP4 was significantly increased in human CRC tissues and correlated with poor prognosis of patients with CRC. Overexpression of Rab11FIP4 in the CRC cell line significantly promoted cell proliferation, migration and invasion in vitro and tumor metastasis in vivo. Furthermore, the results of a coimmunoprecipitation assay and western blot analysis demonstrated that Rab11FIP4 interacted with Rab11 and insulinlike growth factor 1 receptor, and increased the phosphorylation of extracellular signalregulated kinase 1/2 and AKT serine/threonine kinase. In addition, hypoxia contributed to the upregulation of Rab11FIP4 expression via hypoxiainducible factor1α activation of the Rab11FIP4 promoter. In conclusion, the results of the present study suggest that Rab11FIP4 may act as an oncogene in CRC, and may be a potential therapeutic target for the treatment of patients with CRC.
Subject(s)
Carrier Proteins/genetics , Colonic Neoplasms/genetics , Gene Expression Regulation, Neoplastic , Hypoxia/genetics , Insulin-Like Growth Factor I/genetics , Membrane Proteins/genetics , rab GTP-Binding Proteins/genetics , Adaptor Proteins, Signal Transducing/genetics , Adaptor Proteins, Signal Transducing/metabolism , Animals , Carrier Proteins/metabolism , Cell Hypoxia , Cell Movement , Cell Proliferation , Colonic Neoplasms/metabolism , Colonic Neoplasms/mortality , Colonic Neoplasms/pathology , HCT116 Cells , Humans , Hypoxia/metabolism , Hypoxia/mortality , Hypoxia/pathology , Insulin-Like Growth Factor I/metabolism , Male , Membrane Proteins/metabolism , Mice , Mice, Nude , Mitogen-Activated Protein Kinase 1/genetics , Mitogen-Activated Protein Kinase 1/metabolism , Mitogen-Activated Protein Kinase 3/genetics , Mitogen-Activated Protein Kinase 3/metabolism , Neoplasm Transplantation , Prognosis , Signal Transduction , Survival Analysis , rab GTP-Binding Proteins/metabolismABSTRACT
Rab11-family interacting proteins (Rab11-FIPs) belong to an evolutionarily conserved protein family and act as effector molecules for the Rab11 family of small GTPases. Recent evidence suggests that Rab11-FIPs have important roles in tumor progression and metastasis. However, the contribution of Rab11-FIPs to colorectal carcinoma (CRC) remains elusive. Our study focuses on elucidating the role of Rab11-FIP2 in the migration and invasion of colorectal cancer cells. We firstly found upregulation of Rab11-FIP2 in CRC tissues compared with peritumor tissues by oncomine data-mining analysis, western blot analysis and immunohistochemistry (IHC) analysis, respectively. Then, we demonstrated that knockdown of Rab11-FIP2 via siRNAs transfection resulted in a decrease in migration and invasion of CRC cells, while overexpression of Rab11-FIP2 via lentiviral infection increased migration and invasion of CRC cells. In addition, we verified that Rab11-FIP2 promoted migration and invasion of CRC cells through upregulating MMP7 expression. Finally, using several kinase inhibitors, our results showed that Rab11-FIP2 regulated MMP7 expression through activating PI3K/Akt signaling. Our data suggested a potential role of Rab11-FIP2 in tumor progression and provided novel insights into the mechanism of how Rab11-FIP2 positively regulated cell migration and invasion in CRC cells.
Subject(s)
Carrier Proteins/metabolism , Colorectal Neoplasms/metabolism , Colorectal Neoplasms/pathology , Matrix Metalloproteinase 7/metabolism , Membrane Proteins/metabolism , Phosphatidylinositol 3-Kinases/metabolism , Proto-Oncogene Proteins c-akt/metabolism , Cell Line, Tumor , Cell Movement , Gene Expression Regulation, Neoplastic , Humans , Neoplasm Invasiveness , Signal Transduction , rab GTP-Binding ProteinsABSTRACT
OBJECTIVE: To investigate the haemodynamic and respiratory changes following intravenous administration with midazolam, pavulon or both of them in the patients having incoordination between spontaneous breathing and mechanical ventilation. METHODS: Thirty patients having incoordination between spontaneous breathing and mechanical ventilation were randomly assigned to receiving intravenous injection of pavulon (group 1), midazolam (group 3), and both (group 2) respectively with 10 cases in each group. The degree of coordination between spontaneous breathing and mechanical ventilation, blood pressure (BP), heart rate (HR), respiration frequency (RF), oxygen saturation of pulse (SpO(2)) were observed before the medication and at 5, 15, 30 and 60 minutes following the administration of drugs in all the patients. RESULTS: Incoordination between spontaneous breathing and mechanical ventilation, fast RF, decreased SpO(2) were observed before the drug in all patients. Improvement of respiratory was significant in group 2. Patients in group 2 were in excellent coordination between spontaneous breathing and mechanical ventilation, reaching 100% within 30 minutes after administration, and lasting longer. The haemodynamics maintained stable and a significant improvement in respiration and SpO(2) were found. BP and HR were elevated significantly, and RF and hypoxemia were improved, and the degree of coordination between spontaneous breathing and mechanical ventilation reached 100% 5 minutes after the drug, but with shorter duration in group 1. There were no obvious changes in BP, HR, RF and hypoxemia, and the degree of coordination between spontaneous breathing and mechanical ventilation was lowest in group 3. CONCLUSION: The combined use of midazolam and pavulon has little influence on circulation, and it also can maintain the coordination between spontaneous breathing and mechanical ventilation. It is suggested that the combined use of midazolam and pavulon is an optimal way to improve the ventilatory function in mechanical ventilation.
