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Objective To study the role of MDC/CCL22-CCR4 axis in milky spots in peritoneal carcinomatosis of gastric cancer.Methods We examined the expression of CCR4 in 5 gastric cancer cell lines by RT-PCR and Western-blot.Cell growth rate of BGC-823 cell lines before and after incubated with CCL22 was detemined by MTT assay.The effects of CCL22 on invasion of BGC-823 cells into Matrigel were measured using Transwell test.Immunohistochemistry was conducted to detect the expression of CCR4.Results CCR4 mRNA and protein expression were detected in all 5 human gastric cancer cell lines:CCL22had a direct promotive effect on BGC-823 cell prolifetation and invasion.There was CCR4 expression in Stomach neoplasms.The expression of CCR4 in primary tumor was correlated with tumor pathological differentiation(P<0.05),but not with age of patients,gender,location of tumor,tumor size,tumor stage,lymph node metastasis(P>0.05).Conclusions The MDC/CCL22-CCR4 axis may be one of the mechanism of peritoneal metastasis in milky spot in gastric cancer.
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Objective To explore the effects of systemic lupus erythematosus (SLE) susceptibility gene IFIT1 on chemokine expression in RAW264.7 macrophages and its possible role in the pathogenesis of SLE. Methods The expression vector of pEGFP-N1/IFIT1 was transfected into RAW264.7 cells by electroporation. 24 h after transfection, cells were stimulated with LPS ( 1 μg/ml). The transcriptional levels of chemokine MIP-1α, RANTES, CCL9, CXCL2 and IP-10 were measured at various time points after stimu-lation using real-time quantitative PCR. The chemokine expression levels in the kidneys of 8 week-old NZB/NZW F1 mice were also determined by real time PCR. Results Compared with cells transfected with null vector, IFIT1 high RAW264.7 cells produced significantly increased levels of MIP-1α, RANTES, CCL9, CXCL2 and IP-10 both at 4 h and 24 h after stimulation (P<0.05). Chemokine expression levels were signi-ficantly elevated in kidneys of 8 week-old NZB/NZW F1 mice compared with those of 8 week-old BALB/c mice controls (P<0.05). Conclusion IFIT1 may participate in target organ damages in SLE via augmentation of chemokine production by macrophage cells.
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ObjectiveTo investigate the effect of resveratrol on the expressions of E-selectin and monocyte chemoattractant protein-1 (MCP-1) in activated endothelial cells.Methods After being pretreated with resveratrol followed by tumor necrosis factor-α (TNF-α) stimulation, human umbilical vein endothelial cells (HUVEC) were randomly divided into three groups: TNF group,resveratrol+TNF-α group and control group. The expression of E-selectin molecule on the surface of HUVEC was detected by flow cytometric analysis and the mRNA expressions of E-selectin and MCP -1 were determined by semiquantitative RT-PCR. ResultsTNF-α induced the expression of E-selectin and MCP-I of HUVEC.Resveratrol (10 μmol/L) inhibited E-selectin expression.The positive cells of E-selectin in TNF group, resveratrol + TNF-α group and control group were(47.84±3.2)%, (15.3±1.7)% and (3.74±1.6)%, respectively, and the differences among the three groups were statistically significant (P<0.05). Conclusions Resveratrol may contribute to the anti-atherosclerotic effect by inhibiting the expression of E-seleetin and MCP-1 of HUVEC.
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Objectives To develop and evaluate a real-time fluorescent quantitative RT-PCR method for the detection of a novel CXC chemokine macrophage inflammatory protein-2?(MIP-2?)mRNA expression based on TaqMan technique.Method The expression of MIP-2? mRNA was detected by TaqMan real time fluorscet quantitative RT-PCR. Results The dynamic range of the assay varied from 102 to 107 copies. The intra- and inter-assay coefficient variation (CV) were less than
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Objective:To discuss the effect of bone marrow on chemotaxis of aggregation of macrophages in venous thrombosis.Methods:SD adult rats were divided randomly into normal group,sham operation group,thrombus group and rhG-CSF group.Peripheral blood was harvested at 30min before operation and on the 7th and 14th day after operation.Level of MCP-3 in peripheral blood was detected with ELISA.Inferior vena cava specimens of thromus group and rhG-CSF group were collected at postoperative 7th and 14th day.With the application of RT-PCR,expression of MCP-3mRNA in thrombus was detected.Results:The level of MCP-3 in peripheral blood of rhG-CSF group was significantly higher than that before operation and also higher than the levels of MCP-3 in peripheral blood of sham operation group and thrombus group on the 14th day after operation,(P
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AIM: To observe the effects of Egr-1 gene knockout on the expression of inflammatory-related factors in pancreatic tissue in a mouse acute pancreatitis model.METHODS: The experimental pancreatitis was induced by high-dose of cearulein in wildtype mice and Egr-1 knockout mice.The pancreatitis indexes,such as serum amylase,pancreata edema,and myeloperoxidase(MPO) levels in pancreata and lungs were recorded.The mRNA levels of tissue factor(TF),plasminogen activator inhibitor(PAI-1),monocyte chemoattractant protein(MCP-1),Gro-1,IL-6 and ICAM-1 were measured by quantitative PCR.RESULTS: Contrary to wildtype mice,typical pancreatitis was not induced by high-dose cearulein in the Egr-1 knockout mice,not only markedly reduced edema in pancreata and lungs,but decreased MPO levels in lungs as well were found.Furthermore,the mRNA of TF,PAI,MCAP,ICAM-1 and IL-6 in pancreata were significantly decreased in Egr-1 knockout mice.CONCLUSION: The severity of pancreatitis and lung damage is ameliorated in Egr-1 knockout mice stimulated by high-dosage of cearulein,which was probably mediated by decreasing expression of inflammatory-related factors in pancreata,such as TF,PAI,MCP-1,ICAM-1 and IL-6.