Procaine regulates CXCR7 and affects AKT/STAT3 signaling pathway to inhibit viability,migration and invasion of bladder cancer cells

AIM To investigate the effects of procaine (PCA) and CXC chemokine receptor 7 (CXCR7) on the viability, migration and invasion of bladder cancer cells and its potential mechanism. METHODS Human bladder cancer RT4 cells were treated with PCA at different concentrations, and were divided into PBS group (without PCA treatment), PCA group (treated with 4 mmol/L PCA), siRNA negative control (si-Con) group (transfected with si-Con), CX?CR7 siRNA (si-CXCR7) group (transfected with si-CXCR7), PCA+pcDNA group (treated with 4 mmol/L PCA and transfected with pcDNA) and PCA+pcDNA-CXCR7 group (treated with 4 mmol/L PCA and transfected with pcDNA-CX?CR7). The siRNA and pcDNA were transfected into the RT4 cells by liposome method. The mRNA expression of CX?CR7 in the RT4 cells was detected by RT-qPCR. The cell viability was measured by CCK-8 assay. The invasion and migration abilities of the cells were detected by Transwell assays. The protein levels of CXCR7, AKT, STAT3, p-AKT and p-STAT3 were determined by Western blot . RESULTS Compared with PBS group, the viability, migration ability and invasion ability of the RT4 cells treated with PCA at different concentrations were significantly decreased (P<0.05), and the expression of CXCR7 at mRNA and protein levels in PCA group was also significantly decreased (P<0.05). Compared with si-Con group, the expression of CXCR7 at mRNA and protein levels in si-CXCR7 group was significantly decreased, and the viability, migration ability and invasion ability of the cells were also significantly decreased (P<0.05). Compared with PCA+pcDNA group, the expression of CXCR7 at mRNA and protein levels in PCA+pcDNA-CXCR7 group was significantly increased, and the viability, migration ability and invasion ability of the cells were also significantly increased (P<0.05). Compared with PBS group, the protein levels of p-AKT and p-STAT3 in PCA group were significantly decreased(P<0.05). Compared with PCA+pcDNA group, the protein levels of p-AKT and p-STAT3 in PCA+pcDNA-CX?CR7 group were significantly increased (P<0.05). No significant difference in the protein levels of AKT and STAT3 among the groups was observed. CONCLUSION Treatment with PCA inhibits the viability, migration and invasion of bladder cancer cells by inhibiting the expression of CXCR7. Over-expression of CXCR7 reverses this effect of PCA. Its mechanism may be related to AKT/STAT3 signaling pathway.     

Effects of marrow stromal cell-mediated microenvironment on human lung adenocarcinoma A549 cells

AIM: To investigate the effects of marrow stromal cell line HS-5 on human lung adenocarcinoma A549 cells in the tumor microenvironment. METHODS: The effects of HS-5 cell-conditioned medium (HS-5-CM) on the viability and migration ability of A549 cells were detected by MTT assay and wound-healing assay. After treatment with HS-5-CM, the expression of CX3C chemokine receptor 1 (CX3CR1) at mRNA level in the A549 cells was examined by qPCR. The protein levels of p-ERK and ERK in the A549 cells treated with MAPK/ERK pathway inhibitor U0126 were observed by Western blot, the migration ability of the A549 cells was measured by wound-healing assay, and the protein expression of CX3CR1 was determined by Western blot. RESULTS: HS-5-CM promoted the viability and migration ability of the A549 cells (P<0.01). The expression of CX3CR1 at mRNA level in the A549 cells was increased after treatment with HS-5-CM. MAPK/ERK inhibitor U0126 inhibited the activation of MAPK/ERK signaling pathway (P<0.01), and reduced the migration ability (P<0.01) and the expression of CX3CR1 (P<0.05) in the A549 cells. CONCLUSION: HS-5-CM significantly promotes the A549 cell viability and migration ability. Activation of MAPK/ERK signaling pathway and the expression of CX3CR1 may play a important role in this process.     

Regulatory effect of astragaloside IV on SDF-1α/CXCR4 in EPCs

AIM: To investigate the effects of astragaloside IV (AS-IV) on chemokine receptor 4 (CXCR4) and stromal cell-derived factor 1α (SDF-1α) in endothelial progenitor cells (EPCs) and its mechanism. METHODS: Rat bone marrow-derived EPCs were cultured in vitro. The proliferation, adhesion, migration, apoptosis and tube formation capacity of EPCs treated with AS-IV and AMD3100, a specific blocker of CXCR4, were observed. The effects of AS-IV on the expression of SDF-1α/CXCR4 at mRNA and protein levels and the protein level of p-CXCR4 in the EPCs were determined. RESULTS: AS-IV significantly enhanced the proliferation, adhesion, migration and tube formation abilities of EPCs, reduced the apoptosis of EPCs, and up-regulated the mRNA and protein expression of SDF-1α and CXCR4 and the p-CXCR4 protein level in the EPCs. On the other hand, AMD3100 blocked the up-regulating effect of AS-IV on the mRNA and protein expression of CXCR4 and the p-CXCR4 protein level in the EPCs, but did not affect the effect of AS-IV on the expression of SDF-1α. CONCLUSION: AS-IV might enhance the biological function of EPCs by regulating the expression of SDF-1α/CXCR in EPCs.     

Molecular mechanism of BMSC intracerebral transplantation in improving learning and memory abilities of AD mice

AIM: To investigate the effect of bone marrow mesenchymal stem cell (BMSC) transplantation on learning and memory abilities and pathological changes of Alzheimer disease (AD) mice and the molecular mechanisms. METHODS: C57/BL6 wild-type (WT) and transgenic (Tg) mice were randomly divided into 4 groups:WT/PBS group, WT/BMSCs group, Tg/PBS group and Tg/BMSCs group. The mice were administered with PBS or BMSCs via intracerebroventricular injection. Spatial learning and memory abilities of the mice were evaluated by Morris water maze test on the 3rd day after surgery. Real-time PCR was applied to detect the mRNA expression of CX3C chemokine ligand 1 (CX3CL1), CX3C chemokine receptor 1 (CX3CR1), IL-1β, TNF-α, Nurr1, YM1, insulin-degrading enzyme (IDE) and matrix metalloproteinase 9 (MMP9). The protein levels of CX3CL1 and Aβ42 were measured by ELISA. Western blot was used to detect the protein expression of postsynaptic density protein 95 (PSD95) and synaptophysin (SYP). RESULTS: The transplanted BMSCs were observed near the hippocampus of APP/PS1 mice on the 10th postoperative day. The escape latency of the mice in Tg/PBS group was significantly longer than that in the WT/PBS mice (P<0.05). Compared with Tg/PBS group, the escape latency of Tg/BMSCs group was significantly shorter (P<0.05), and the mRNA and protein levels of CX3CL1 in Tg/BMSCs group were significantly higher than those in Tg/PBS group (P<0.01). The results of immunohistofluorescence staining showed that BMSC transplantation promoted the activation of microglia in the brain of WT and Tg mice. The mRNA expression of YM1 was up-regulated in WT/BMSCs group and Tg/BMSCs group (P<0.05). Compared with WT/PBS mice, the mRNA expression of TNF-α in the cortex and hippocampus of Tg/PBS group was significantly increased (P<0.05), and the mRNA expression of Nurr1 in the cortex was significantly decreased (P<0.01). Meanwhile, the mRNA expression of TNF-α in the cortex of Tg/BMSCs mice was decreased (P<0.01) and the mRNA expression of CX3CR1 and Nurr1 was up-regulated compared with Tg/PBS group (P<0.05). The results of Western blot showed that the protein levels of PSD95, p85, p110 and p-Akt in Tg/BMSCs group were significantly higher than those in Tg/PBS group (P<0.05). Finally, BMSC transplantation reduced the protein level of Aβ42 in APP/PS1 mice (P<0.05), and increased the mRNA expression of IDE and MMP9 in the hippocampus (P<0.05). CONCLUSION: BMSC transplantation modulates neuroinflammatory responses and promotes neuroprotective factor and synaptic protein expression, thus improving the learning and memory abilities in the APP/PS1 mice, which may be achieved by up-regulating the expression of CX3CL1.     

Effects of marrow stromal cell-mediated microenvironment on human hepatocellular carcinoma cell line SMMC-7721

AIM: To investigate the effects of marrow stromal HS-5 cells on hepatocellular carcinoma SMMC-7721 cells in the tumor microenvironment. METHODS: The effects of HS-5 cell-conditioned medium (HS-5-CM) on the proliferation, migration and invasion abilities of SMMC-7721 cells were detected by MTT, wound-healing and Transwell assays. After co-culture of SMMC-7721 cells with HS-5 cells in the Transwell chamber, the expression of chemokine CCL5 and its receptor CCR5 at mRNA and protein levels in SMMC-7721 cells was examined by quantitative real-time PCR (qRT-PCR), ELISA or Western blotting. Akt and p-Akt473 protein levels in SMMC-7721 cells treated with PI3K inhibitor LY294002 were observed by Western blotting. RESULTS: HS-5-CM promoted the proliferation, migration and invasion abilities of SMMC-7721 cells. The expression of CCL5 and CCR5 at mRNA and protein levels in SMMC-7721 cells was increased after co-cultured with HS-5 cells. PI3K inhibitor LY294002 inhibited the activation of PI3K-Akt signaling pathway and the secretion of CCL5 in SMMC-7721 cells after co-cultured with HS-5 cells. CONCLUSION: HS-5 cells significantly promote the proliferation, migration and invasion abilities of SMMC-7721 cells. Co-culture of SMMC-7721 cells with HS-5 cells activates PI3K-Akt signaling pathway to increase the secretion of CCL5 in SMMC-7721 cells.     

CCL3 promotes hBMSC proliferation and inhibits exosome secretion

AIM: To explore the regulatory effect of chemokine CCL3 on exosome secretion from human bone marrow mesenchymal stem cells (hBMSCs).METHODS: hBMSCs were stimulated with chemokine CCL3 at different concentrations in vitro. The proliferation of hBMSCs was measured by CCK-8 assay and viable cell counting. Exosome secretion from hBMSCs was qualitatively analyzed by transmission electron microscope (TEM) and flow cytometry, and the quantitative analysis was carried out by flow cytometry and nanoparticle tracking analysis (NTA).RESULTS: Compared with control group, the viability of the hBMSCs detected by CCK-8 assay was increased when hBMSCs were treated with CCL3 (P<0.05). The results of viable cell counting demonstrated that the number of hBMSCs was raised in CCL3 group in a dose-dependent manner (P<0.05). The results of flow cytometry showed that hBMSCs expressed 3 CCL3-related specific receptors, CCR1, CCR5 and CCR9. Compared with control group, the fluorescence intensity of CCR9 in CCL3 group was obviously enhanced. However, no significant difference of fluorescence intensity for CCR5 and CCR1 was observed between the 2 groups. The results of NTA demonstrated that the secretion capacity of CCL3-induced hBMSCs was far less than that in control group (P<0.05). However, the microvesicles larger than 100 nm in CCL3 groups were increased (P<0.05). The above results indicated that the higher concentration of CCL3 induced the lower secretion of exosomes. In addition, the results of flow cytometry demonstrated that CCL3-induced hBMSCs showed lower quantity of CD9⁺ exosomes than those in control group (P<0.01).CONCLUSION: CCL3 promotes the proliferation of hBMSCs but depresses the secretion of exosomes in a dose-dependent manner. CCL3 affects the size distribution of exosomes and increases the number of nonfunctional microvesicles of larger than 100 nm in size. CCL3 induces the expression of CCR9 in hBMSCs.     

Expression of fractalkine and CX3CR1 in renal tissues of patients with WHO class IV lupus nephritis

AIM: To observe the expression of fractalkine, and its receptor, CX3CR1, in renal tissues of patients with diffuse proliferative lupus glomerulonephritis (WHO class IV), minimal glomerular abnormalities, and normal kidney. Meanwhile, the correlation among the expression of fractalkine, CX3CR1 and CD68-positive macrophages was investigated, and the role of fractalkine and CX3CR1 in the pathogenesis of lupus nephritis was discussed. METHODS: The expressions of fractalkine, CX3CR1 and CD68 were detected immunohistochemically in kidney tissue sections obtained from twenty-one patients with WHO class IV lupus nephritis, eighteen cases with minimal glomerular abnormalities, and eight normal kidneys which were no abnormality under light microscope. RESULTS: (1) Fractalkine was generally indistinguishable in tissue sections from normal kidney and minimal glomerular abnormalities. CX3CR1-positive cells and CD68-positive macrophages were sparsely detected in the glomeruli and in the cortical interstitium. (2) There were considerable CX3CR1-positive cells and macrophages in both the glomeruli and the interstitium in sections from class IV lupus nephritis. The number of CX3CR1-positive cells significantly correlated with the number of macrophages in the glomeruli and in the interstitium respectively (r=0.956, P<0.01 and r=0.965, P<0.01). (3) Significant expression of fractalkine was seen in the cortical renal tubules from class IV lupus nephritis. The percentage of fractalkine-positive tubules significantly correlated with the number of CX3CR1-positive cells and macrophages in the interstitium respectively (r=0.720, P<0.01 and r=0.770, P<0.01). CONCLUSION: These expression patterns show that fractalkine and CX3CR1 may play an important role in the pathogenesis of class IV lupus nephritis.     

Role of CXCR4 in changes of protein C system in ulcerative colitis mice

AIM: To explore the role of chemokine receptor CXCR4 in the pathogenesis of protein C system (PCS) in ulcerative colitis (UC).METHODS: In vivo, the mice were divided into control group and UC group. The macroscopic score, microscopic score and ulcer index were assessed. The mRNA levels and activity of myeloperoxidase (MPO), cyclooxygenase-2 (COX-2), stromal cell-derived factor-1α (SDF-1α) and monocyte chemotactic protein 1 (MCP-1) both in colonic tissue and plasma were determined. The expression and location of CXCR4, β-arrestin, p-JNK, endothelial cell protein C receptor (EPCR) and thrombomodulin (TM) were detected. The activity of protein C (PC) and protein S (PS) was measured in each group. In vitro, mouse colonic microvascular endothelial cells were isolated, cultured and identified. Both CXCR4-overexpressing and CXCR4-silencing colonic mucosa microvascular endothelial cells were constructed. The effects of SDF-1α on the protein levels of EPCR, TM, β-arrestin and p-JNK, and on the activity of PC, PS and activated protein C (APC) were observed.RESULTS: Compared with control group, UC mice showed increased gross score, histopathological score and ulcer index (P<0.05). The mRNA levels and activity of MPO, COX-2, SDF-1α and MCP-1 in colon and plasma were increased (P<0.01). The protein levels of CXCR4, β-arrestin and p-JNK were up-regulated, EPCR expression was down-regulated in colon, and the activity of PC and PS in plasma was decreased (P<0.05 or P<0.01). CXCR4 overexpression further aggravated SDF-1α-induced PCS inhibition in colonic mucosa microvascular endothelial cells, and further up-regulated the protein levels of β-arrestin and p-JNK (P<0.05).CONCLUSION: PCS is inhibited in UC. CXCR4 is involved in the regulation of PCS inhibition by mediating chemokines and acting on colonic mucosa microvascular endothelial cells through β-arrestin-JNK pathway.     

Fractalkine and CX3CR1 have intimate relationships with chemotaxis and adhesion of leukocyte

Fractalkine has been recognized as the first member of the fourth chemokine family recently and its receptor, CX3CR1, was ident ified afterwards. This pair of ligand/receptor play important roles in the migration and adhesion of peripheral leukocyte through different ways. They are also likely to be related to the embryogenesis and regeneration of several organs and may possess some functions in the central nervous system.     

Immunologic mechanism of CXCL10 and its receptor involved in endometriosis

AIM: To investigate the immunologic mechanism of CXC chemokine ligand 10(CXCL10) and its receptor CXC chemokine receptor 3(CXCR3) involved in the process of endometriosis (EM). METHODS: Serum samples were collected from 3 groups: EM patients without operation (n=76), EM patients with operation (n=10) and the normal control persons (n=76). CXCL10 and CA125 concentrations were detected by means of ELISA and chemiluminometry. Cell surface antigens on the activated PBMC-CD3 and CXCR3, as well as CXCR3 subgene-CXCR3A and CXCR3B were tested by flow cytometry (FC) and RT-PCR when PBMC was separated from women with EM (n=10) and without EM (n=10), and then activated. RESULTS: Serum CXCL10 concentrations between three groups were significanly different (P<0.05). Compared to normal control group, although the supernatant CXCL10 concentration and CD3+/CXCR3+PBMC number in EM group has no significant difference (P>0.05), highly expressed CXCR3B in EM group rather than CXCR3A was observed. CONCLUSION: CXCL10 in women with EM is low, indicating that it plays a vital role in the process of EM and immune system of the women with EM is defected and impaired. The immunoreactivity of PBMC from both EM patients and normal person is same to activated signal, but the productions are different: PBMC in EM group mainly express CXCR3B but PBMC in normal person mainly express CXCR3A after activation, which may be one of the immune mechanisms that EM escapes from immunological lethal effect of the infected host.     

CX3CR1 mediates the neuroprotective effect of triptolide on 1-methyl-4-phenylpyridinium-induced hemiparkinson rats

AIM: To investigate the effect of triptolide on the inhibition of microglial activation in 1-methyl-4-phenyl pyridinium (MPP⁺)-induced hemiparkinson disease rats.METHODS: The rat model of Parkinson disease was established by intranigral injection of MPP⁺. The rats were randomly divided into sham group, MPP⁺ group, triptolide group and vehicle group. The survival of dopaminergic neurons was detected by the immunofluorescence of tyrosine hydroxylase (TH) in the substantia nigra (SN). The activation of microglia was determined by immunofluorescence of OX-42 (microglia marker) in the SN. The expression of chemokine receptor CX3CR1 in SN was measured by Western blotting.RESULTS: Intranigral injection of MPP⁺ increased the fluorescence intensity of the microglial marker, and promoted DA neuron degenerative death. Immunohistological analysis showed that the OX-42 density was decreased (P<0.01) and tyrosine hydroxylase (TH) positive neurons were increased in the triptolide group (P<0.01). The expression of CX3CR1 was lower in triptolide group than that in model group (P<0.05).CONCLUSION: Triptolide may improve PA neurons function in MPP⁺-induced rats through inhibiting CX3CR1 expression and microglial activation.     

Effects of erythropoietin on expression of SDF-1 and its receptor in peripheral blood endothelial progenitor cells from rats with chronic renal failure

AIM:To investigate the effects of erythropoietin (EPO) on the expression of homing factors in peripheral blood endothelial progenitor cells (EPCs) from rats with chronic renal failure (CRF). METHODS:The CRF model was established by a two-stage 5/6 nephrectomy procedure in rats. Experimental rats were randomly divided into three groups: sham operation group, CRF model group and EPO treatment group. From the third week after the second stage of 5/6 nephrectomy procedure, rats in EPO treatment group received subcutaneous injection of human recombinant EPO at 50 U/kg every time and three times a week for 6 weeks, and then all the rats were sacrificed. EPCs were isolated from rat peripheral blood and primarily cultured. The mobilization, angiogenesis and functional activity of EPCs in vitro were detected. The mRNA and protein expression of EPO, EPO receptor (EPOR), stromal cell-derived factor 1 (SDF-1) and CXC chemokine receptor 4 (CXCR4) in EPCs was also detected by the methods of real-time PCR and Western blotting. RESULTS:Compared with CRF model group, the expression of EPO and EPOR in EPCs in EPO treatment group was significantly up-regulated (P<0.05). Moreover, the expression of SDF-1 and its receptor CXCR4 in EPCs was also up-regulated by administration of EPO (P<0.05). CONCLUSION: EPO can mobilize EPCs from CRF rat peripheral blood, which may be associated with the increased expression of SDF-1 and its receptor CXCR4.     

Effects of propofol on expression of CXCR4/CXCR7 and migration ability of human breast cancer MCF-7 cells in vitro

AIM:To investigate the effects of propofol on the expression of CXC-chemokine receptor (CXCR)4/CXCR7 and migration ability of human breast cancer MCF-7 cells in vitro. METHODS:MCF-7 cells were randomly divided into 4 groups,control group, lipid emulsion group, 3 mg/L and 8 mg/L propofol group. The cell viability was measured by MTT assay. The migration ability was detected by wound-healing assay and Transwell assay. The mRNA level of CXCR4/CXCR7 was detected by RT-qPCR. The protein expression leve of CXCR4/CXCR7 was determined by Western blot. RESULTS:Compared with control group, the scratching healing rates in 3 and 8 mg/L propofol group were decreased (P<0.05), and the chemotactic index was also decreased (P<0.05). The protein expression level of CXCR4/CXCR7 was decreased in 3 and 8 mg/L propofol group(P<0.05). However, both the mRNA level of CXCR4/CXCR7 and the viability of the MCF-7 cells kept no change. CONCLUSION:Propofol down-regulates the protein expression of CXCR4/CXCR7 and inhibits the migration ability of breast cancer MCF-7 cells in vitro.     

Specific surface markers of Th1/Th2 subsets and their identification

CD4+Tcells can be divided into Th1/Th2 subsets.Th1/Th2 imbalance participates many disease processes.A stable surface marker distinguishing Th1 and Th2 will greatly facilitate the investigation of Th1/Th2 interaction.Several surface molecules have been reported to be differentialy expressed between Th1 and Th2 cells.LAG-3, active ligands for P-and E-selectin, IL-18R, IL-12Rβ2, CC chemokine receptor(CCR5)were shown to be dominantly expressed on Th1 cells, whereas expression of CD30, ST2L, CRTH2, CCR3, CCR4 was reported to be preferential to Th2 cells.In this review, several surface molecules were mainly discussed.     

Effect of pregnancy uterine microenvironment on the expression of NKG2A,NKG2D and their ligands in decidual NK cells

AIM：To investigate the expression of NKG2A,NKG2D and their ligands in pregnancy uterine micro-environment and to probe the function of NKG2A and NKG2D imbalance expression during the immunotolerance at the fetal-maternal boundary.METHODS：Decidual lymphocytes and peripheral lymphocytes were obtained from 30 women during 6-9 weeks of pregnancy who were undergoing selective termination.FACS technology was used to detect NK cells number and NKG2A,NKG2D expression.RT-PCR was used to investigate HLA-E and MICA mRNA in trophoblast tissue.RESULTS：Natural killer cells predominate,accounting for 70% of pregnancy endometrial lymphocytes.FACS results indicated that NKG2A was significantly increased in decidual NK cells as compared with that in peripheral NK cells,accounting for 97.86%±1.75% and 33.35%±10.92%.The difference between them in NKG2A expression was significant (P<0.05).Although the levels of NKG2D in decidual NK cells were similar to that in peripheral NK cells,accounting for 93.21%±4.52% and 97.80%±1.72%,respectively,the difference between them in NKG2A expression was also significant (P<0.05).At the same time,we found HLA-E mRNA in the trophoblast tissue.There was no proof yet on the expression of NKG2D ligand-MICA on trophoblast.CONCLUSIONS：Natural killer cells are the predominant lymphocytes during the first trimester of decidual tissue.These cells have a great different antigenic phenotype with peripheral natural killer cells.Decidual natural killer cells have high expression of NKG2A and trophoblast tissue expressed HLA-E.This may be an important factor contributing to the immunotolerance at the fetal - maternal boundary during pregnancy.     

Resveratrol modulates SDF-1/CXCR4 signaling to inhibit hypoxia-ischemia-induced apoptosis of neurons from the brain of newborn rats

AIM:To investigate the effect of resveratrol (RSV) on apoptosis and stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) pathway in hypoxia-ischemia-induced neurons from the brain of newborn rats and its mechanism. METHODS:The cortex neurons from the brain of newborn rats were given oxygen-glucose deprivation (OGD) treatment to mimic neonatal hypoxic-ischemic encephalopathy (HIE). The cortex neurons were randomly divided into 4 groups:control group, HIE model group, HIE+RSV-low (10 μmol/L) group, and HIE+RSV-high (50 μmol/L) group. After OGD treatment for 2 h, the neurons were cultured with indicated dose of RSV for 24 h. The apoptosis was analyzed by flow cytometry. Western blot was used to determine the levels of apoptosis-related proteins, SDF-1 and CXCR4. Real-time PCR was used to detect the mRNA expression of SDF-1 and CXCR4. Additionally, to explore the effects of RSV on cell apoptosis and apoptosis-related proteins after the suppression of SDF-1/CXCR4 signaling, a CXCR4 antagonist AMD3100, and RSV were used to co-treat OGD-injured neurons for 24 h. RESULTS:RSV alleviated OGD-induced neuronal apoptosis, down-regulated cleaved caspase-3 and cytochrome C levels, and up-regulated the ratio of Bcl-2/Bax. Compared with the control group, OGD treatment increased the expression of SDF-1 and CXCR4 (P<0.05). Compared with the HIE model group, RSV further up-regulated the expression of SDF-1 and CXCR4 (P<0.05). AMD3100 reversed the effects of RSV on OGD-induced cell apoptosis. CONCLUSION:RSV suppresses hypoxia-ischemia-induced apoptosis of neurons from the brain of newborn rats via up-regulating SDF-1/CXCR4 signaling pathway.     

MCP-1/CCR2 signaling pathway mediates alcohol-induced angiogenesis in breast cancer

AIM To investigate the role of monocyte chemoattractant protein-1 (MCP-1) and its receptor CC chemokine receptor 2 (CCR2) in ethanol-promoted breast cancer angiogenesis and the underlying mechanism. METH?ODS: A mouse model of transplanted breast tumor with moderate alcohol consumption was established. The correlations between the expression of MCP-1/CCR2 and the expression of angiogenesis markers [platelet endothelial cell adhesion molecule-1 (PECAM-1) and vascular endothelial growth factor (VEGF)] in tumor tissues were examined by immunohistochemistry. In vitro, a 3D tumor-endothelial co-culture system was established to observe tumor angiogenesis and the role of MCP-1/CCR2 signaling pathway in alcohol-mediated angiogenesis. The cell migration ability was detected to clarify whether MCP-1/CCR2 enhanced cell mobility to form new vessels. RESULTS MCP-1 and CCR2 were both highly expressed in the breast tumor tissues of tumor-bearing mice consuming alcohol, and their expression levels were consistent with the angiogenic markers PECAM-1 and VEGF (P<0.05). The interaction between mouse breast cancer E0771 cells and endothelial cells was observed to promote angiogenesis in the 3D tumor-endothelial co-culture system with or without alcohol stimulation. MCP-1 promoted this kind of tumor angiogenesis, while CCR2 antagonist effectively inhibited the tumor angiogenesis and especially blocked alcohol-induced angiogenesis. Activation of MCP-1/CCR2 signaling pathway enhanced the migration ability of endothelial cells. CONCLUSION The MCP-1/CCR2 signaling pathway plays an important role in promoting the angiogenesis of breast cancer stimulated by alcohol. The mechanism might be that MCP-1 improves the migration of endothelial cells and then promotes angiogenesis.