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OBJECTIVE: Cyclophilin A (CyPA) is secreted under inflammatory conditions by various cell types. Whereas the important role of intracellular CyPA for platelet function has been reported, the effect of extracellular CyPA on platelet function has not been investigated yet. APPROACH AND RESULTS: Inhibition of extracellular CyPA through a novel specific inhibitor MM284 reduced thrombus after ferric chloride-induced injury in vivo. In vitro extracellular CyPA enhanced thrombus formation even in CyPA(-/-) platelets. Treatment of isolated platelets with recombinant CyPA resulted in platelet degranulation in a time- and dose-dependent manner. Inhibition of the platelet surface receptor extracellular matrix metalloproteinase inducer (cluster of differentiation 147) by an anticluster of differentiation 147 monoclonal antibody significantly reduced CyPA-dependent platelet degranulation. Pretreatment of platelets with CyPA enhanced their recruitment to mouse carotid arteries after arterial injury, which could be inhibited by an anticluster of differentiation 147 monoclonal antibody (intravital microscopy). The role of extracellular CyPA in adhesion could be confirmed by infusing CyPA(-/-) platelets in CyPA(+/+) mice and by infusing CyPA(+/+) platelets in CyPA(-/-) mice. Stimulation of platelets with CyPA induced phosphorylation of Akt, which could in turn be inhibited in the presence of phosphoinositid-3-kinase inhibitors. Akt-1(-/-) platelets revealed a markedly decreased degranulation on CyPA stimulation. Finally, ADP-induced platelet aggregation was attenuated by MM284, as well as by inhibiting paracrine-secreted CyPA without directly affecting Ca(2+)-signaling. CONCLUSIONS: Extracellular CyPA activates platelets via cluster of differentiation 147-mediated phosphoinositid-3-kinase/Akt-signaling, leading to enhanced adhesion and thrombus formation independently of intracellular CyPA. Targeting extracellular CyPA via a specific inhibitor may be a promising strategy for platelet inhibition without affecting critical functions of intracellular CyPA.
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Basigina/sangue , Plaquetas/enzimologia , Ciclofilina A/sangue , Fosfatidilinositol 3-Quinases/sangue , Adesividade Plaquetária , Proteínas Proto-Oncogênicas c-akt/sangue , Transdução de Sinais , Trombose/enzimologia , Animais , Plaquetas/efeitos dos fármacos , Lesões das Artérias Carótidas/sangue , Lesões das Artérias Carótidas/enzimologia , Lesões das Artérias Carótidas/genética , Degranulação Celular/efeitos dos fármacos , Cloretos , Ciclofilina A/antagonistas & inibidores , Ciclofilina A/genética , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Compostos Férricos , Fibrinolíticos/farmacologia , Humanos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Adesividade Plaquetária/efeitos dos fármacos , Agregação Plaquetária/efeitos dos fármacos , Inibidores da Agregação Plaquetária/farmacologia , Proteínas Proto-Oncogênicas c-akt/deficiência , Proteínas Proto-Oncogênicas c-akt/genética , Transdução de Sinais/efeitos dos fármacos , Trombose/sangue , Trombose/induzido quimicamente , Trombose/genética , Trombose/prevenção & controle , Fatores de TempoRESUMO
BACKGROUND: Matrix metalloproteinases (MMPs) play a key role in inflammatory periodontal disease. Synergistically enhanced MMP-2 expression in a coculture of human gingival fibroblasts (HGFs) and human monocytic U937 cells was observed. Crosstalk between these two cells via the extracellular matrix metalloproteinase inducer (EMMPRIN) was demonstrated. METHODS: Enzyme levels of MMP-2 in HGFs and direct coculture with U937 were examined by zymography. MMP-2 and EMMPRIN expressions of HGFs and U937 were determined in coculture and conditioned cultures (using supernatants from HGF- or U937-conditioned medium). The crosstalk was evaluated by EMMPRIN extrasupplement and EMMPRIN inhibition, through pretreatment of U937 with cyclosporine-A. RESULTS: Direct coculturing of HGFs and U937 enhanced MMP-2 enzyme level and mRNA expression. Coculturing also increased membranous EMMPRIN expression of U937, but not from HGFs. In conditioned cultures, mRNA expression of MMP-2 increased in HGFs which received U937-conditioned medium. Increased MMP-2 was not observed in U937 with HGF-conditioned medium, although mRNA expression of EMMPRIN increased. Enhanced MMP-2 was observed after administration of exogenous EMMPRIN in HGFs; however, reduced MMP-2 enzyme level was noted if EMMPRIN of cocultured U937 was inhibited. CONCLUSIONS: In the coculture of HGFs and U937, upregulated EMMPRIN expression in U937, which may be triggered by HGFs, can enhance MMP-2 expression in HGFs. Crosstalk between HGFs and U937 involving MMP-2 from HGFs was proposed; EMMPRIN from U937 may play a particular role.
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Basigina/fisiologia , Gengiva/metabolismo , Metaloproteinase 2 da Matriz/metabolismo , Fibroblastos , Humanos , Células U937RESUMO
Objective To investigate the correlation between the expression of CD147 and MUC5ac in peripheral small cell lung cancer and the computed tomography (CT) signs.Methods Forty two cases of peripheral non-small cell lung cancer (NSCLC) confirmed by operation and pathology were examined by CT.The positive expression of CD147 and MUC5ac was detected by immunohistochemical staining and the relationship between the expression of CD147 and MUC5ac was analyzed.Results CD147 and MUC5ac were closely related to the lymphatic metastasis of peripheral non-small cell lung cancer (P < 0.05).The positive expression of CD147 is significantly related to tumor size,deep lobulation,spinous process and lymphadenopathy (P < 0.05).MUC5ac is significantly related to tumor size,deep lobulation,spiculation,vascular convergence sign,extrapleural fat Line disappearance,and mediastinal lymphadenopathy (P <0.05).Conclusions These results indicate that CD147 and MUC5ac play an important role in the invasion and metastasis of peripheral lung cancer.
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Objective To investigate the expressions of extracellular matrix metalloproteinase inducer molecule CD147 and epidermal growth factor receptor (EGFR) protein in gastric cancer and their relationship with the pathological behavior and prognosis.Methods Immunohistochemical staining was used to detect the expressions of CD147 and EGFR protein in 63 cases of gastric cancer and 40 cases of paracancerous normal tissues.The study was also combined with analysis of the pathological behavior and clinical follow-up survey of gastric cancer.Results The positive expression rates of CD147 and EGFR protein in gastric cancer were 77.8% (49/63) and 42.9% (27/63),respectively,which were higher than those in paracancerous normal tissues 25.0% and 0 (x2 =27.851,P =0.000;x2 =23.233,P =0.000).In gastric cancer,the expressions of CD147 and EGFR were closely related to the depth invasion (x2 =5.992,P =0.014;x2 =13.148,P =0.000),TNM stages (.x2 =8.481,P=0.004;x2=14.589,P=0.000),lymph node metastasis (x2 =5.270,P=0.022;x2=20.261,P =0.000) and prognosis of patients (x2 =5.606,P =0.018;x2 =4.982,P =0.026).The expression of CD147 was also significantly correlated with Ki-67 expression (x2 =10.481,P =0.001).CD147 expression was positively correlated with EGFR expression(r =0.309,P =0.014).Conclusion The expressions of CD147 and EGFR are closely related to carcinogenesis,metastasis and survival time of gastric cancer.Detecting the expressions of CD147 and EGFR may be prognostic indicators of gastric cancer.
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[ ABSTRACT] AIM: To study the autophagy of prostate cancer PC-3 cells induced by CD147 in vitro.ME-THODS:Themethod of amino acid starvation to induce autophagy was used.The expression of CD147 was detected by Western blotting.To study the functional effects of CD147 on autophagy in prostate cancer PC-3 cells, the down-regulation of CD147 expression was induced by the technique of RNAi.The conversion of autophagic marker protein LC3-I to LC3-II was determined by Western blotting.The cell death after starvation-induced autophagy was analyzed by trypan blue exclu-sion assay.RESULTS:The CD147 expression gradually increased in starvation-induced autophagy.The down-regulation of CD147 significantly increased the expression of autophagy-related protein LC3-II compared with control group.Mean-while, the cell death rates increased from (19.3 ±3.1)%and (22.3 ±3.5)%in control groups to (38.4 ±3.1)%in si-lencing the expression of CD147 in the PC-3 cells (P<0.05).CONCLUSION:CD147 inhibits starvation-induced auto-phgy and autophagy death in the prostate cancer PC-3 cells.
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Objective To investigate the effects of the leflunomide active metabolite (A771726) on the expression of phorbol-12-myristate-13-acetate (PMA) -induced CD147, matrix metallo-proteinase (MMP)-2 and MMP-9 on THP-1 cells. Methods THP-1 cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum. For all experiments, THP-1 cells were cultured at an initial density of 5×105/ml. Before A771726 treatment, cells were cultured with serum-free RPMI-1640 medium for 12 h, THP-1cells were co-cultured with PMA at three different concentrations of A771726 (5, 15 , 45 μg/ml) for 24 h.The mRNA expression of CD147, MMP-2 and MMP-9 was measured by real-time PCR. CD147 expression on the cells were evaluated by flow cytometric analysis. The activity of MMP-2 and MMP-9 were evaluated by gelatin zymography. Statistical differences among the groups were tested by one-way ANOVA or KruskalWallis test. Results The expression of CD147, MMP-2 and MMP-9 were upgraded by the PMA. The expression of CD147 on THP-1 cells was inhibited significantly by A771726 in a dose-dependent pattern (P<0.01). The mean fluorescence intensity (MFI) of CD147 in positive control group was 109.5±3.8, the MFI in A771726 (5, 15, 45 μg/ml) group were 73.3±2.5, 64.5±2.3, 40.9±2.7, respectively. The expression of MMP-2, MMP-9 mRNA and the activity of MMP-2, MMP-9 in the supernatant was inhibited significantly by A771726 (P<0.01). The expression of CD147 mRNA was not inhibited significantly by A771726 (P>0.05).Conclusion Leflunomide active metabolite (A771726) can inhibit the expression of PMA-induced CD147,MMP-2 and MMP-9 on THP-1 Cells.
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Objective:To study the expression of extracellular matrix metalloproteinase (MMP-9) inducer CD147 and MMP-9 protein in hepatocellular carcinoma (HCC) and explore their correlation with invasion, metastasis, and prognosis of HCC.Methods:SP immunohistochemistry was used to evaluate the expression of CD147 and MMP-9 in 83 HCC tissues and 15 normal liver tissues. Results:The positive rate was 73.5% for MMP-9 and 68.7% for CD147, significantly higher than normal tissues (MMP-9: 6.7%;CD147: 0%, P<0.05). Expressions of CD147 and MMP-9 were positively correlated with the tumor size, pTNM stages, differentiation degree, the portal vein cancer embolism, and postoperative recurrence (P<0.05), but not with the patient's age, gender, the serum level of alpha-fetal protein (AFP), and the Child-Pugh grades. Patients with positive expression of CD147 and MMP-9 had shorter survival time than those with negative expression of CD147 and MMP-9. Expression of CD147 was positively related with expression of MMP-9 in HCC tissues.Conclusion:CD147 and MMP-9 had positive correlation with tumor size, pTNM stages, differentiation degree, portal vein cancer embolism, and postoperative recurrence. Detection of the expression of CD147 and MMP-9 may be helpful to make a comprehensive prediction of the prognosis of HCC.
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Objective To investigate the expression of extracellular matrix metalloproteinase inducer (CD147), nuclear factor kappa B (NF-κB) in esophageal squamous cell carcinoma tissue and probe their relationship to invasion and metastasis of esophageal carcinoma. Methods Immunochemical staining was performed to detect the expression of CD147 and NF-κB protein in 40 cases of esophageal carcinoma tissues and 40 cases of normal control. Results The positive rate of CD147 and NF-κB in esophageal carcinoma tissues and in adjacent noncancerous tissues were 77.5%/87.5% Vs 25%/42. 5%, P<0.05.The positive rate of their prophase and metaphase-advanced stage esophageal carcinoma tissues were 33.3%/90.3% Vs 55.6%/96.8%, P<0.05..The positive rate of non-metastatic and metastatic esophageal carcinoma tissues were 28.6%/87.9% Vs 42.9%/97%, P<0.05.Conclusions Our results suggested that the increased expression of CD147 and NF-κB contributed to tumor angiogenesis in esophageal squamous cell carcinoma, which might be through the common pathway.
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Objective To investigate the effects of small interfering RNA(siRNA)on the expression of CD147 on peripheral blood T-lymphocytes from patients with psoriasis vulgaris,and its effect on the proliferation and activation of these cells.Methods Peripheral blood monouclear cells(PBMC)were obtained from 10 patients with psoriasis vulgaris,and T lymphocytes were isolated.CD147 siRNA was chemically synthesized,then electroporated into the peripheral T-lymphocytes.Untransfected cells,blank-transfected cells and unspecifically transfected cells served as the control.After 24-,48-,72-and 96-hour incubation,RT-PCR was used to detect the mRNA expression of CD147 in these cells.MTT assay and flow cytometry were utilized to assess the proliferation of these cellas,and the expression Of CD25 at 24,48,and 72 hours after the transfection.Results Compared with untransfected cells,the mRNA expression of CD147 declined significantly in CD147 siRNA-transfected cells at 24 hours(P<0.05),reached to the minimum at 48 hours (P<0.01):there was no significant difierence in the expression of CD147 between the two groups of cells at 96 hours after the transfection(P>0.05).There was a decrease of cell proliferation level by(44.5±3.13)%,(50.7±3.5)%and(53.98±4.15)%in CD147 siRNA-transfected cells 24,48 and 72 hours following the transfection,respectively;the corresponding decrease in blank-transfected cells was (37.28±3.56)%,(33.73±3.29)%,and(28.80±1.49)%,respectively,and that in unspecifically transfected cells,(31.29±2.46)%,(36.1±2.62)%and(32.08±2.78)%,respectively.A significant decrease was observed in the proliferation of CD147 siRNA-transfected cells compared with that of blank-transfected cells and unspecifically transfected cells at these three time points(P<0.05,0.01,0.01 respectively).The expression rate of CD25 at 24,48 and 72 hours was(47.23±3.65)%,(31.50±4.22)%and(23.05±4.15)%,respectively,on CD147 siRNA-transfected cells,and,(80.2±4.8)%,(81.6±3.35)%and(83.5±4.1)%,respectively,on untransfeeted cells;the differences between the two groups at the three time points were statistically significant(all P<0.01).Conclusion CD147 is correlated with cell proliferation and activation of peripheral T lymphoeytes from patients with psoriasis vulgaris,and may serve as a new treatment target for psoriasis.
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Objective To investigate the expression and role of CD147in the differentiation of nor-mal keratinocytes and squamous cell carcinoma(SCC)cells.Methods CD147expression was analyzed immunohistochemically in20cases of verruca vulgaris(VV),various benign,premalignant and malignant epidermal tumors including21cases of seborrheic keratosis(SK),20actinic keratosis(AK),20Bowen' s disease(BD)and57squamous cell carcinoma.SCCs were classified using Broder's system of grading.Im-munoblot analysis was used to observe CD147expression in normal keratinocyte(HaCaT)and SCC cell(HSC-5)in vitro during their differentiation process induced by calcium.The effects of high-calcium medi-um culture and CD147antibody on the differentiation-related morphology of HaCaT and HSC-5cells were also observed.Results The same staining pattern of CD147was shown in20VV and21SK specimens as in normal epidermis.Positive CD147staining throughout the lesion was shown in4of20AK and7of20BD specimens.Positive CD147staining at the periphery of tumor nests was shown in8of20gradeⅠSCC specimens.CD147expressed throughout tumor nests in16of20gradeⅡSCC and all of the17gradeⅢSCC specimens.Immunoblot analysis revealed decreased CD147expression in both HaCaT and HSC-5cells during differentiation process induced by calcium.Not only high-calcium medium but also CD147antibody could induce differentiation-related morphology of both HaCaT and HSC-5cells in vitro.Conclusion These results suggest that CD147is a novel low-differentiation marker of keratinocyte,which might inhibit the dif-ferentiation of both normal keratinocyte and SCC cell.
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Objective To study the expression and pathologic significance of CD147,cyclophilin A (CyPA)and cyclophilin B(CyPB)in psoriatic lesions.Methods Immunohistochemical method was ap- plied to detect the expression of CD147,CyPA and CyPB in skin specimens of 15 patients with psoriasis pustulosa(PP),20 patients with progressive psoriasis vulgaris(PPV),and 20 patients with inactive psori- asis vulgaris(IPV).Immunoreactivity intensity distribution index(IRIDI)was calculated to assess the expres- sion intensity of CD147,CyPA and CyPB.Results CD147,CyPA and CyPB were detected in all speci- mens.The IRIDI scores of CD147 and CyPA were significantly higher in keratinocytes of psoriatic lesions than in those of the control specimens(all P0.05).The IRIDI score of CyPB in T lymphocytes of PP lesions was significantly elevated than that in PPV lesions,which was in turn higher than that in IPV lesions(all P0.05).Conclusion CD147,CyPA and CyPB may play a role in the occurrence and development of psoriasis.
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Objective To investigate the expressions and significances of extracellular matrix metalloproteinase inducer (EMMPRIN),the matrix metalloproteinases-2,-9 (MMP-2,-9),and the vascular endothelial growth factor (VEGF) in endometriosis (EM). Methods The expressions of the EMMPRIN,MMP-2,MMP-9 and VEGF were detected by SABC immunohistochemistry in 57 tissue samples of ectopic endometrium and 51 samples of eutopic endometrium obtained from 57 EM patients,and in 48 normal endometrium from patients with non-endometriosis. Correlation analysis was carried out on the expressions of these proteins and pathologic staging. Results The EMMPRIN,MMP-2,MMP-9 and VEGF in ectopic endometrium were expressed at significantly higher levels (0.723?0.114,0.729?0.109,0.692?0.089,0.704?0.103,respectively) than those in eutopic endometrium group (0.396?0.097,0.365?0.023,0.349?0.030,0.370?0.129,respectively,P