Effect Of Monocyte Chemotactic Protein-1 Research Articles

Monocyte chemotactic protein-1 deficiency reduces spontaneous metastasis of Lewis lung carcinoma in mice fed a high-fat diet

Adipose-produced pro-inflammatory cytokines contribute to obesity and cancer. This 2×2 experiment was designed to investigate effects of monocyte chemotactic protein-1 (MCP-1) deficiency on pulmonary metastasis of Lewis lung carcinoma (LLC) in MCP-1 deficient and wild-type mice fed a modified AIN93G diet containing 16% and 45% of energy from corn oil, respectively. The high-fat diet significantly increased the number and size (cross-sectional area and volume) of lung metastases compared to the AIN93G control diet. Deficiency in MCP-1 reduced lung metastases by 37% in high-fat diet-fed mice; it reduced metastatic cross-sectional area by 46% and volume by 69% compared to wild-type mice. Adipose and plasma concentrations of MCP-1 were significantly higher in high-fat diet-fed wild-type mice than in their AIN93G-fed counterparts; they were not detectable in MCP-1 deficient mice regardless of diet. Plasma concentrations of plasminogen activator inhibitor-1, tumor necrosis factor-α, vascular endothelial growth factor and tissue inhibitor of metalloproteinase-1 were significantly higher in MCP-1 deficient mice compared to wild-type mice. We conclude that adipose-produced MCP-1 contributes to high-fat diet-enhanced metastasis. While MCP-1 deficiency reduces metastasis, the elevation of pro-inflammatory cytokines and angiogenic factors in the absence of MCP-1 may support the metastatic development and growth of LLC in MCP-1 deficient mice.

MCPIP1 mediates silica-induced cell migration in human pulmonary fibroblasts.

Silicosis is a systemic disease caused by inhaling silicon dioxide (SiO2). Phagocytosis of SiO2 in the lungs initiates an inflammatory cascade that results in fibroblast proliferation and migration followed by fibrosis. According to previous data from our laboratory, monocyte chemotactic protein-1 (MCP-1) plays a critical role in fibroblast proliferation and migration in conventional two-dimensional (2D) monolayer cultures. The present study aimed to explore the downstream cascade of MCP-1 in both 2D and three-dimensional (3D) cell culture models of silicosis. Experiments using primary cultured adult human pulmonary fibroblasts (HPF-a) demonstrated the following: 1) SiO2 treatment induces expression of MCP-1-induced protein (MCPIP1) in a time- and dose-dependent manner in both 2D and 3D cultures; 2) the MAPK and phosphatidylinositol-3-kinase (PI3K)/Akt pathways are involved in SiO2-induced MCPIP1 expression; and 3) MCPIP1 induction mediates the SiO2-induced increase in cell migration in both 2D and 3D cultures. The effect of MCP-1 in silicosis occurs mainly through MCPIP1, which, in turn, mediates the observed SiO2-induced increase in pulmonary fibroblast migration. However, the time frame for MCPIP1 induction differed between 2D and 3D cultures, indicating that, compared with conventional 2D cell culture systems, 3D culture may be useful for analyses of fibroblast physiology under conditions that more closely resemble in vivo environments. Our study determined the link between fibroblast-derived MCPIP1 and SiO2-induced cell migration, and this finding provides novel evidence of the potential of MCPIP1 in the development of novel therapeutic strategies for silicosis.

Effects of Monocyte Chemotactic Protein-1 and Nuclear Factor of Kappa B Pathway in Rejection of Cardiac Allograft in Rat

BackgroundGraft rejection is a key obstacle to successful heart transplantation. We sought to investigate the expression and role of monocyte chemotactic protein-1 (MCP-1) in rejection of cardiac allografts, as well as the role of the nuclear factor of kappa B (NF-κB) pathway. MethodsHeterotopic cervical heart transplantation was performed using a modified cuff-technique. Recipient rats (n = 12) underwent acute rejection (AR) without any treatment (AR group). The remaining rats (3 groups; n = 12/group) were treated with Cyclosporine A (CsA; CsA group), immunologic tolerance (IT; IT group), and pyrrolidine dithiocarbamate (PDTC; NF-κB inhibitor; PDTC group). We studied the inflammatory infiltration and myocardial fibrosis of cardiac allografts with hematoxylin-eosin (HE) and Masson staining, and detected the expression of MCP-1 by immunohistochemistry and Western blotting. Cardiac allograft vasculopathy (CAV) also was evaluated using van Gieson staining. ResultsThe survival time of the PDTC group (142.37 ± 24.28 days) was significantly longer than that of the AR group (6.54 ± 2.47 days; P = .00073) and the CsA group (93.51 ± 20.17 days; P = .0052). Myocardial fibrosis and CAV in the PDTC group were significantly attenuated compared with the CsA group (P < .01). The expression of MCP-1 in the IT group was markedly lower than in the other 3 groups (P < .05). The expression of MCP-1 in the PDTC group was also significantly lower than the CsA group (1.15 ± 0.27 vs 1.58 ± 0.17; P = .016). ConclusionThese findings suggest that the expression level of MCP-1 could be monitored to reflect the severity of cardiac allograft rejection. PDTC can significantly prevent the rejection of cardiac allografts by inhibiting MCP-1 expression via the suppression of the NF-κB pathway.

MCP/CCR2 signaling is essential for recruitment of mesenchymal progenitor cells during the early phase of fracture healing.

ObjectiveThe purpose of this study was to investigate chemokine profiles and their functional roles in the early phase of fracture healing in mouse models.MethodsThe expression profiles of chemokines were examined during fracture healing in wild-type (WT) mice using a polymerase chain reaction array and histological staining. The functional effect of monocyte chemotactic protein-1 (MCP-1) on primary mouse bone marrow stromal cells (mBMSCs) was evaluated using an in vitro migration assay. MCP-1−/− and C-C chemokine receptor 2 (CCR2)−/− mice were fractured and evaluated by histological staining and micro-computed tomography (micro-CT). RS102895, an antagonist of CCR2, was continuously administered in WT mice before or after rib fracture and evaluated by histological staining and micro-CT. Bone graft exchange models were created in WT and MCP-1−/− mice and were evaluated by histological staining and micro-CT.Results MCP-1 and MCP-3 expression in the early phase of fracture healing were up-regulated, and high levels of MCP-1 and MCP-3 protein expression observed in the periosteum and endosteum in the same period. MCP-1, but not MCP-3, increased migration of mBMSCs in a dose-dependent manner. Fracture healing in MCP-1−/− and CCR2−/− mice was delayed compared with WT mice on day 21. Administration of RS102895 in the early, but not in the late phase, caused delayed fracture healing. Transplantation of WT-derived graft into host MCP-1−/− mice significantly increased new bone formation in the bone graft exchange models. Furthermore, marked induction of MCP-1 expression in the periosteum and endosteum was observed around the WT-derived graft in the host MCP-1−/− mouse. Conversely, transplantation of MCP-1−/− mouse-derived grafts into host WT mice markedly decreased new bone formation.ConclusionsMCP-1/CCR2 signaling in the periosteum and endosteum is essential for the recruitment of mesenchymal progenitor cells in the early phase of fracture healing.

Association of Polymorphisms inMCP-1, CCR2,andCCR5Genes with the Risk and Clinicopathological Characteristics of Prostate Cancer

The aim of our study was to determine the effect of monocyte chemotactic protein-1 (MCP-1), CC chemokine receptor 2 (CCR2), and CC chemokine receptor 5 (CCR5) gene polymorphisms on the susceptibility and clinicopathological characteristics of prostate cancer. Genotyping was performed by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) method in 156 histopathologically confirmed prostate cancer patients and 152 healthy subjects. Individuals with AA genotype or at least one A allele of CCR2 V64I gene polymorphism had a higher risk for prostate cancer as compared with those with GG genotype (p=0.010 and p=0.028, respectively). CCR5 Δ32/wt genotype and CCR5 Δ32 allele were also found to be involved in the susceptibility to prostate cancer (p=0.028 and p=0.030, respectively). However, there was no significant association between MCP-1-2518 A/G gene polymorphism and prostate cancer risk. Prostate cancer patients carrying AA genotype or at least one A allele of CCR2 V64I had significantly increased risk for high stage disease (p=0.002 and p=0.039, respectively) and metastasis (p=0.004 and p=0.022, respectively). The CCR2 A allele (64I allele) was significantly associated with high T stage (p=0.001) and metastasis (p=0.005) as compared with CCR2 G allele (64V allele). Our data indicate that gene polymorphism of CCR2 V64I may influence the susceptibility and clinicopathological characteristics of prostate cancer and CCR5 Δ32 allele may also be an important risk factor for prostate cancer in Turkish men population.

Effects of Peroxisome Proliferator-Activated Receptor (PPAR) Delta on the Growth and Invasion of a Thyroid Cancer Cell Line

Background: Peroxisome proliferator-activated receptor delta (PPAR-δ) is a ligand-activated nuclear transcription factor that is associated with many diseases, such as diabetes, obesity, metabolic syndrome, and cancer. However, the function of PPAR-δ is controversial in carcinogenesis since its ligands may inhibit or promote the growth of cancer cells. The purpose of this study was to determine the effect of GW501516, the specific agonist of PPAR-δ, in the growth and invasiveness of thyroid cancer cell lines by modulation of the target genes, ANGPTL-4 and MCP-1. Methods: Three kinds of human cancer cell lines, FRO (thyroid anaplastic carcinoma), NPA (melanoma), and ARO (colon cancer) were treated with GW501516 in serum-free media. Cell viability was assayed using a colorimetric cell counting kit-8 assay. The changes in the level of expression of PPAR-δ and its target genes, angiopoietin-like protein-4 (ANGPTL-4) and monocyte chemotactic protein-1 (MCP-1), were determined by RT-PCR analysis and invasiveness was assessed by a cell invasion assay kit. Results: GW501516 inhibited the cell growth of cancer cell lines in a dose-dependent manner and modulated the stimulation of ANGPTL-4, as well as inhibition of MCP-1. These effects were more prominent in NPA and ARO, but less effective in the thyroid cancer cell l ine, which had higher PPAR-δ and lower ANGPTL-4 mRNA levels. The inhibitory effects of GW501516 on cancer invasiveness had a similar pattern. Conclusion: The activation of PPAR-δ by GW501516 reduced the cell growth and invasiveness of the thyroid cancer cell line. This effect of GW501516 was associated with a stimulatory effect of ANGPTL4 and an inhibitory effect of MCP-1 in cancer cell lines. GW501516 was less effective in the thyroid cancer cell line, which had a low basal ANGPTL-4 mRNA level. The findings of our study serve as an impetus for further studies to elucidate the precise role of ANGPTL-4 and PPAR-δ in carcinogenesis. (J Korean Endocr

Expression of monocyte chemotactic protein-1 in peripheral blood mononuclear cells of children with Kawasaki disease and its relation to coronary artery impairment

Kawasaki disease (KD) is a febrile illness of childhood. The etiology of KD remains unknown. Multiple theories exist, including an infectious etiology and an immunological abnormality. Cardiac involvement ranges from myocarditis and pericarditis in the acute stage to the development of coronary artery aneurysms later in the course. The present study aimed to explore the effect of monocyte chemotactic protein-1 (MCP-1) in Kawasaki disease and its relationship with damage to the coronary arteries during the development of KD. Plasma MCP-1 concentrations were measured by enzyme-linked immunosorbent assay (ELISA), and MCP-1 mRNA expression in peripheral blood mononuclear cells (PBMC) was measured by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) in comparison of three groups: 56 patients with KD, 60 age-matched patients with non-infectious diseases, and 66 age-matched febrile patients with various diseases. Plasma MCP-1 concentration and MCP-1 mRNA expression in PBMC of patients with active KD [(409.55 +/- 97.42) pg/ml] and (1.97 +/- 0.77) were higher than those of control group. Plasma MCP-1 levels and MCP-1 mRNA expression of inactive KD group [(301.64 +/- 71.55) pg/ml] and (1.31 +/- 0.39) were significantly higher than those of non-infectious diseases patients. There was a marked increase in patients with inactive KD than those of non-infective patients, but there were no significant differences between inactive KD and febrile patients. Plasma MCP-1 levels and MCP-1 mRNA expression were markedly increased in KD patients with coronary artery lesions than those in patients without coronary artery lesions. Plasma MCP-1 concentration and MCP-1 mRNA expression in PBMC were significantly increased in patients with KD, and they were higher in KD with coronary artery lesions. It indicates that MCP-1 may be a useful parameter for monitoring disease activity in patients with KD.

Migration of retinal pigment epithelial cells in vitro modulated by monocyte chemotactic protein-1: enhancement and inhibition.

The migration of retinal pigment epithelial (RPE) cells is an initial step in the development of proliferative vitreoretinopathy (PVR). This in vitro study was carried out to investigate the effects of monocyte chemotactic protein-1 (MCP-1) on the migration and proliferation of RPE cells. We used an in vitro wound healing model in which a small area of a confluent monolayer of human RPE (HRPE) cells was denuded with a razor blade. The cultures were subsequently incubated with MCP-1, IL-1beta, TNF-alpha, or combinations thereof. Neutralizing IgG1 of antihuman MCP-1, dexamethasone (DEX) or daunorubicin were also added to the cultures to test their inhibitory effects on migration of RPE cells. HRPE migration was measured as the number of cells that entered the denuded area. The effect of MCP-1 on proliferation of HRPE cells was examined by MTT assay. MCP-1 stimulated HRPE cell migration in a dose-dependent manner. IL-1beta or TNF-alpha slightly stimulated HRPE cell migration, but adding anti-MCP- IgG1 significantly reduced this effect. MCP-1-induced migration could be inhibited by DEX but not by daunorubicin. MCP-1 did not show a significant effect on HRPE cell proliferation. MCP-1 stimulates HRPE cell migration, suggesting that this chemokine regulates the development of PVR at the initial stage. The migration of HRPE cells induced by IL-1beta and TNF-alpha may be associated with the MCP-1 that HRPE cells secretes under the stimulation of these two cytokines. The knowledge that MCP-1-induced migration of HRPE cells is inhibited by DEX may be useful in devising an effective treatment for PVR.

Expression of MacMARCKS restores cell adhesion to ICAM-1-coated surface.

To evaluate the role of MacMARCKS, a major substrate of protein kinase C, in cell adhesion, we selected a macrophage cell line, Wehi 274.1.7. Although surface expression of beta2-integrins can be detected on these cells, they lack the phorbol ester- or chemokine-induced adhesion to ICAM-1-coated surface, an event mediated by beta2-integrins. Concomitantly, these cells lack expression of both MacMARCKS and its homologue, MARCKS. When wild type MacMARCKS was expressed in these cells, the phorbol ester-induced adhesion to ICAM-1-coated surface increased approximately 5-fold compared to vector transfected control cells. To further investigate the potential physiological role of MacMARCKS in this adhesion event, we also tested the effect of monocyte chemotactic protein-1, and a 3-fold increase in the adhesion to ICAM-1-coated surface was observed with MacMARCKS-transfected cells. Therefore, these data suggest that MacMARCKS is an essential component in regulating cell adhesion.

Expression and Hormonal Regulation of Monocyte Chemotactic Protein-1 in Myometrium and Leiomyomata

Objective: The pathogenesis of leiomyoma likely involves interactions of sex steroids with paracrine growth factors or cytokines resulting in modulation of local immunity. Monocyte chemotactic protein-1 (MCP-1) is a chemotactic and activating factor for monocytes and is produced by multiple tumors and has antitumor effects. We investigated the expression of MCP-1 in leiomyoma and myometrium as well as the regulatory role of steroid hormones and cytokines on MCP-1 expression and the effect of MCP-1 on the proliferation of leiomyoma cells.Design: Prospective study.Setting: University medical center.Patient(s): Women with (n = 20) or without (n = 11) leiomyoma.Intervention(s): First, MCP-1 messenger RNA (mRNA) levels in myometrium and leiomyoma were measured, and then myometrial and leiomyoma cells in culture were treated with steroid hormones and cytokines.Main Outcome Measure(s): The MCP-1 mRNA was evaluated by Northern analysis. Immunoreactive MCP-1 in cell cultures was quantified by ELISA. Leiomyoma cell proliferation was assessed with [3H]thymidine incorporation.Result(s): The MCP-1 mRNA levels in myometrial samples were 4.7-fold higher than in the leiomyoma samples. Myometrial MCP-1 mRNA levels were 2.4-fold higher in secretory than in proliferative phase samples. The highest MCP-1 levels were observed in samples from women using GnRH analogues. Estradiol and progestins, alone or in combination, resulted in a decrease in MCP-1 protein production. There was an increase in the proliferation of leiomyoma cells treated with anti-MCP-1 neutralizing antibody.Conclusion(s): These findings suggest that MCP-1 may have antineoplastic activity in leiomyomata and that sex steroids may be exerting their growth stimulatory effect in leiomyomata through down-regulation of MCP-1.

Costimulation of Fibroblast Collagen and Transforming Growth Factor β1 Gene Expression by Monocyte Chemoattractant Protein-1 via Specific Receptors

Recent studies indicate potential roles of monocyte chemotactic protein-1 (MCP-1) in recruitment of monocytes to sites of inflammation. However, their increased expression does not always correlate with monocyte influx, suggesting other possible biological activities for this member of the C-C chemokine family. In view of its potential role in regulating extracellular matrix expression in fibrotic disorders, the effects of MCP-1 on lung fibroblast collagen expression were evaluated. Isolated rat lung fibroblasts were treated with increasing doses of MCP-1 for variable periods of time and examined for effects on collagen synthesis and expression of procollagen alpha1(I) mRNA expression. The results show that MCP-1 was able to stimulate collagen expression in these cells in a dose-dependent manner but required over 24 h for significant elevation to occur. In view of this delayed time course, the possibility of mediation via endogenous transforming growth factor beta (TGFbeta) was tested by the ability of anti-TGFbeta antibody to inhibit this MCP-1 stimulation of collagen expression. Significant but incomplete inhibition by this antibody was observed. Pretreatment of the cells with antisense but not by sense or missense TGFbeta1 oligodeoxyribonucleotides caused essentially complete inhibition of this MCP-1 stimulatory effect. Furthermore, MCP-1 treatment was found to also stimulate TGFbeta secretion and mRNA expression, which was also abolished by pretreatment with antisense TGFbeta1 oligodeoxyribonucleotides. The kinetics of TGFbeta expression indicates that significant increase preceded that for collagen expression. Binding studies using 125I-labeled MCP-1 indicated the presence of specific and saturable binding sites with a dissociation constant consistent with the dose response curves for stimulation of fibroblast collagen synthesis and TGFbeta activity by MCP-1. These results taken together suggest that MCP-1 stimulates fibroblast collagen expression via specific receptors and endogenous up-regulation of TGFbeta expression. The latter then results in autocrine and/or juxtacrine stimulation of collagen gene expression.

Monocyte Chemotactic Protein-1 Inhibits the Induction of Nitric Oxide Synthase in J774 Cells

We evaluated the effect of monocyte chemotactic protein-1 (MCP-1) on the induction of nitric oxide synthase activity in J774 cells. MCP-l was able to inhibit the production of nitric oxide induced by LPS and IFN-γ in a dose-dependent manner. Moreover, the inhibition was only achieved when the cells were pretrated with MCP-1. No inhibition was observed when MCP-1 was added after stimulation with LPS and IFN-γ. These results demostrate that MCP-1 is able to inhibit the induction of nitric oxide synthesis,