Co-expression of CD147 (EMMPRIN), CD44v3-10, MDR1 and monocarboxylate transporters is associated with prostate cancer drug resistance and progression

Background: The aim of this study is to seek an association between markers of metastatic potential, drug resistance-related protein and monocarboxylate transporters in prostate cancer (CaP). Methods: We evaluated the expression of invasive markers (CD147, CD44v3-10), drug-resistance protein (MDR1) and monocarboxylate transporters (MCT1 and MCT4) in CaP metastatic cell lines and CaP tissue microarrays (n=140) by immunostaining. The co-expression of CD147 and CD44v3-10 with that of MDR1, MCT1 and MCT4 in CaP cell lines was evaluated using confocal microscopy. The relationship between the expression of CD147 and CD44v3-10 and the sensitivity (IC50) to docetaxel in CaP cell lines was assessed using MTT assay. The relationship between expression of CD44v3-10, MDR1 and MCT4 and various clinicopathological CaP progression parameters was examined. Results: CD147 and CD44v3-10 were co-expressed with MDR1, MCT1 and MCT4 in primary and metastatic CaP cells. Both CD147 and CD44v3-10 expression levels were inversely related to docetaxel sensitivity (IC50) in metastatic CaP cell lines. Overexpression of CD44v3-10, MDR1 and MCT4 was found in most primary CaP tissues, and was significantly associated with CaP progression. Conclusions: Our results suggest that the overexpression of CD147, CD44v3-10, MDR1 and MCT4 is associated with CaP progression. Expression of both CD147 and CD44v3-10 is correlated with drug resistance during CaP metastasis and could be a useful potential therapeutic target in advanced disease.

Disrupting the EMMPRIN (CD147)-cyclophilin A interaction reduces infarct size and preserves systolic function after myocardial ischemia and reperfusion.

Objective-Inflammation and proteolysis crucially contribute to myocardial ischemia and reperfusion injury. The extracellular matrix metalloproteinase inducer EMMPRIN (CD147) and its ligand cyclophilin A (CyPA) may be involved in both processes. The aim of the study was to characterize the role of the CD147 and CyPA interplay in myocardial ischemia/reperfusion (I/R) injury.Methods and Results-Immunohistochemistry showed enhanced expression of CD147 and CyPA in myocardial sections from human autopsies of patients who had died from acute myocardial infarction and from mice at 24 hours after I/R. At 24 hours and 7 days after I/R, the infarct size was reduced in CD147(+/-) mice vs CD147(+/+) mice (C57Bl/6), in mice (C57Bl/6) treated with monoclonal antibody anti-CD147 vs control monoclonal antibody, and in CyPA(-/-) mice vs CyPA(+/+) mice (129S6/SvEv), all of which are associated with reduced monocyte and neutrophil recruitment at 24 hours and with a preserved systolic function at 7 days. The combination of CyPA(-/-) mice with anti-CD147 treatment did not yield further protection compared with either inhibition strategy alone. In vitro, treatment with CyPA induced monocyte chemotaxis in a CD147-and phosphatidylinositol 3-kinase-dependent manner and induced monocyte rolling and adhesion to endothelium (human umbilical vein endothelial cells) under flow in a CD147-dependent manner.Conclusion-CD147 and its ligand CyPA are inflammatory mediators after myocardial ischemia and reperfusion and represent potential targets to prevent myocardial I/R injury.

Expressão Imuno-histoquímica de COX-2, EMMPRIN, HIF-1α e GLUT-1 no tecido gengival normal e inflamado

The aim of this study was determine whether an association exists in the gum tissue between the expression of markers of tissue hypoxia (HIF-1α and GLUT-1) with a marker of inflammatory activity (COX-2) and a marker of collagen degradation (EMMPRIN). Was performed immunohistochemistry with antibodies specific for these markers on 60 samples of gingival tissue divided into two groups: gums (n = 26) and gingivitis (n = 34) and expression was analyzed in the epithelial tissue and connective tissue . The reactivity epithelial for COX-2 was observed in only two cases as the HIF-1α, GLUT-1 and EMMPRIN was strongly expressed in the epithelial basal layer and the immunostaining was gradually decreased as the cells away from this layer, and negative in the region suprabasal in most specimens. In connective tissue, and HIF-1α EMMPRIN were strongly positive for most cases analyzed as GLUT-1 was negative in most cases. Immunostaining for COX-2 showed an association with gingival inflammatory infiltrate. The expression of EMMPRIN, HIF-1α and GLUT-1 in normal gums confirms the physiological role of these markers, however there was no association with tissue inflammation. Given the findings we can conclude that the inflammatory changes installed in frames of chronic gingivitis may not be sufficient to activate the factors of hypoxia to levels that can be quantified by immunohistochemical analysis, in addition, the findings are not conclusive in relationship to involvement of EMMPRIN in the secretion of MMPs to degrade collagen in the frames of gingivitis. We suggest the use of technical analysis and quantification of RNA of EMMPRIN and MMPs in order to determine whether collagen degradation observed in gingivitis suffers or not, significant influence of EMMPRIN for secretion and activation of MMPs

Análise da expressão imuno-histoquímica da ciclofilina A, EMMPRIN e MMP-7 na doença periodontal

Periodontal disease is an infectious disease resulting from the immunoinflammatory response of the host to microorganisms present in the dental biofilm which causes tissue destruction. The objective of this study was to evaluate the immunohistochemical expression of cyclophilin A (CYPA), extracellular matrix metalloproteinase inducer (EMMPRIN) and matrix metalloproteinase 7 (MMP-7) in human specimens of clinically healthy gingiva (n=32), biofilm-induced gingivitis (n=28), and chronic periodontitis (n=30). Immunopositivity for CYPA, EMMPRIN and MMP-7 differed significantly between the three groups, with higher percentages of staining in chronic periodontitis specimens, followed by chronic gingivitis and healthy gingiva specimens (p < 0.001). Immunoexpression of CYPA and MMP-7 was higher in the intense inflammatory infiltrate observed mainly in cases of periodontitis. Analysis of possible correlations between the immunoexpression of EMMPRIN, MMP-7 and CYPA and between the expression of these proteins and clinical parameters (probing depth and clinical attachment loss) showed a positive correlation of CYPA expression with MMP-7 (r = 0.831; p < 0.001) and EMMPRIN (r = 0.289; p = 0.006). In addition, there was a significant positive correlation between probing depth and expression of MMP-7 (r = 0.726; p < 0.001), EMMPRIN (r = 0.345; p = 0.001), and CYPA (r = 0.803; p < 0.001). These results suggest that CYPA, EMMPRIN and MMP-7 are associated with the pathogenesis and progression of periodontal disease

Porphyromonas gingivalis-mediated shedding of extracellular matrix metalloproteinase inducer (EMMPRIN) by oral epithelial cells: a potential role in inflammatory periodontal disease

Extracellular matrix metalloproteinase inducer (EMMPRIN) or CD 147 is a transmembrane glycoprotein expressed by various cell types, including oral epithelial cells. Recent studies have brought evidence that EMMPRIN plays a role in periodontitis. In the present study, we investigated the effect of Porphyromonas gingivalis, a major pathogen in chronic periodontitis, on the shedding of membrane-anchored EMMPRIN and on the expression of the EMMPRIN gene by oral epithelial cells. A potential contribution of shed EMMPRIN to the inflammatory process of periodontitis was analyzed by evaluating the effect of recombinant EMMPRIN on cytokine and matrix metalloproteinase (MMP) secretion by human gingival fibroblasts. ELISA and immunofluorescence analyses revealed that P. gingivalis mediated the shedding of epithelial cell-surface EMMPRIN in a dose- and time-dependent manner. Cysteine proteinase (gingipain)-deficient P. gingivalis mutants were used to demonstrate that both Arg- and Lys-gingipain activities are involved in EMMPRIN shedding. Real-time PCR showed that P. gingivalis had no significant effect on the expression of the EMMPRIN gene in epithelial cells. Recombinant EMMPRIN induced the secretion of IL-6 and MMP-3 by gingival fibroblasts, a phenomenon that appears to involve mitogen activated protein kinases. The present study brought to light a new mechanism by which P. gingivalis can promote the inflammatory response during periodontitis. (C) 2011 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

Deteccão de diferentes microorganismos na periodontite crônica, da glicoproteína EMMPRIN (CD-147) e sua correlação com a produção de MMP-2 e MMP-9

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Cinética de expressão e localização de indutor de metaloproteínases (EMMPRIN) durante o desenvolvimento de lesão periapical induzida em camundongos

Pós-graduação em Odontologia - FOAR

Association of matrix metalloproteinase inducer (EMMPRIN) with the expression of matrix metalloproteinases-1, -2 and -9 during periapical lesion development

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Detecção de Periodontopatógenos, Glicoproteína Emmprin (Cd-147) e sua correlação com MMP-2 e MMP-9

Periodontitis is an infectious disease characterized by the secretion of a variety of inflammatory mediators that lead to destruction of tooth supporting tissues, including the possible loss of alveolar bone, in association with infection with multiple species of bacteria. It is estimated that more than 400 species colonize the biofilm and some oral species related to periodontal disease is present in the subgingival including P. gingivalis, T. forsythia and T. denticola. However, other organisms may be related of this disease, as Filifactor allocis and Prevotella tannerae. These microorganisms and subproducts such as endotoxins released into the extracellular lead to the stimulation of metalloproteinase inducer glycoprotein (EMMPRIN, CD-147), which stimulates the release of MMPs by host cells, like fibroblasts and endothelial cells, thus leading to tissue destruction. The objective of this study was to detect F. allocis, P. tannerae, T. denticola and the glycoprotein EMMPRIN (CD-147) and its correlation with MMP-2 and MMP-9 in subgingival fluid samples of patients with chronic periodontitis. Fluids were collected from healthy and disease subgingival sites of 20 healthy individuals before basic periodontal treatment and after of 60 days of treatment. Their DNAs were extracted and portions of the 16S gene were amplified and performed conventional PCR. For immunological analysis and quantification of EMMPRIN (CD-147), MMP-2 and MMP-9 was used ELISA-Sandwich. Results demonstrated that the disease group showed significantly high amounts of T. denticola, F. alocis and P. tannerae when compared with health sites. MMP-2 and MMP-9 were detected in high concentrations with statistically significantly reduction after periodontal treatment to MMP-2, but without correlation with EMMPRIN.

MCT1-, MCT4- ja CD147-proteiinit kehittyvässä kilin pötsissä

Nuoren märehtijän alkaessa syödä kiinteää ravintoa, etumahojen suhteellinen osuus mahoista kasvaa ja niiden seinämän epiteeli alkaa kehittyä mahdollistaakseen ravintoaineiden tehokkaan imeytymisen. Märehtijöillä rehun hiilihydraatit hajoavat pötsissä haihtuviksi rasvahapoiksi, ja monokarboksylaattikuljettajien uskotaan avustavan haihtuvien rasvahappojen imeytymisessä pötsin seinämän läpi. Pötsin seinämässä on todettu olevan ainakin MCT1- ja MCT4 –isoformeja. Nämä tarvitsevat toimiakseen CD147 -proteiinin (myös OX-47, EMMPRIN, HT7 ja basigin), joka on on glykosyloitu integraalinen membraaniproteiini. Tämän tutkimuksen tarkoituksena oli selvittää MCT1-, MCT4- ja CD147 –proteiinien muutoksia pötsin toiminnan kehittymisen aikana. Toisena tavoitteena oli selvittää, voidaanko näytteenä käyttää solukalvojen sijasta pötsin seinämästä tehtyä homogenaattia. Tutkimuksessa käytettiin eri ikäisinä lopetetuista kileistä kerättyjä näytteitä. Kilejä oli yhteensä 31, joista 7 oli 3-21 tunnin ikäisiä, 7 viikon ikäisiä, 7 kahden viikon ikäisiä, 1 kolmen viikon ikäinen, 2 neljän viikon ikäistä, sekä 7 kahdeksan viikon ikäistä. Pötsin seinämästä otettiin näyte, josta valmistettiin homogenaatti ja eristettiin solukalvot eli membraanit. Pötsinäytteistä löydettiin MCT1- ja CD147 –proteiineja, mutta MCT4- isoformia ei ollut havaittavissa. Membraaninäytteissä havaittiin MCT1 -isoformin pitoisuuksien kasvavan iän mukana, paitsi kahdeksan viikon ikäisillä kileillä, joilla MCT1 –isoformin määrät vähenivät merkitsevästi. CD147 –proteiinia oli havaittavissa jo vastasyntyneiden kilien pötsinäytteissä. Membraaninäytteissä CD147 -proteiinin määrä kasvoi lineaarisesti iän mukana ja CD147- proteiinin ja MCT1 –isoformin välillä havaittiin tilastollisesti merkitsevä korrelaatio. Homogenaattinäytteissä MCT1 -isoformin määrissä ei havaittu korrelaatiota iän kanssa. MCT1- ja MCT4 -isoformien solukalvolle siirtymisessä avustavan CD147 –proteiinin ei myöskään havaittu korreloivan koe-eläinten iän tai MCT1 -isoformin kanssa. Membraani- ja homogenaattinäytteistä mitattujen MCT1- ja CD147 -määrien välillä ei ollut korrelaatiota. Haihtuvien rasvahappojen muodostus alkaa, kun eläin aloittaa kiinteän ravinnon syömisen. Tästä seuraa, että haihtuvia rasvahappoja kuljettavia proteiineja tarvitaan epiteelisolujen pinnalle. Tutkimuksessa havaittiin haihtuvia rasvahappoja kuljettavan MCT1 –proteiinin ja sen apuproteiinien määrän lisääntyminen iän myötä. N. 8-11 viikon iässä, jolloin pötsin toiminta on kehittynyt aikuisen eläimen tasolle, MCT1 –proteiinin määrä oli merkitsevästi vähäisempi kuin 4 viikon iässä. Tulosten perusteella homogenaatti ei ole hyvä tapa mitata membraaniproteiinien määrää.

Estudo imuno-histoquímico da presença de miofibroblastos e da expressão do fator transformador de crescimento-beta1, interferon gama, metaloproteinase de matriz 13 e indutor de metaloproteinases de matriz em lesões odontogênicas epiteliais

Myofibroblasts are cells that exhibit a hybrid phenotype, sharing the morphological characteristics of fibroblasts and smooth muscle cells, which is acquired during a process called differentiation. These cells then start to express -SMA, a marker that can be used for their identification. Studies suggest that myofibroblasts are related to the aggressiveness of different tumors and that TGF-1 and IFN- play a role in myofibroblast differentiation, stimulating or inhibiting this differentiation, respectively. The objective of this study was to investigate the role of myofibroblasts in epithelial odontogenic tumors, correlating the presence of these cells with the aggressiveness of the tumor. Immunohistochemistry was used to evaluate the expression of TGF-1 and IFN- in myofibroblast differentiation, as well as the expression of MMP-13, which is activated by myofibroblasts, and of EMMPRIN (extracellular matrix metalloproteinase inducer) as a precursor of this MMP. The sample consisted of 20 solid ameloblastomas, 10 unicystic ameloblastomas, 20 odontogenic keratocysts, and 20 adenomatoid odontogenic tumors. For evaluation of myofibroblasts, anti- -SMA-immunoreactive cells were quantified in connective tissue close to the epithelium. Immunoexpression of TGF-1, IFN-, MMP-13 and EMMPRIN was evaluated in the epithelial and connective tissue components, attributing scores of 0 to 4. The results showed a higher concentration of myofibroblasts in solid ameloblastomas (mean of 30.55), followed by odontogenic keratocysts (22.50), unicystic ameloblastomas (20.80), and adenomatoid odontogenic tumors (19.15) (p=0.001). No significant correlation between TGF-1 and IFN- was observed during the process of myofibroblast differentiation. There was also no correlation between the quantity of myofibroblasts and MMP-13 expression. Significant correlations were found between MMP-13 and TGF-1 (r=0.087; p=0.011), between MMP- 13 and IFN- (r=0.348; p=0.003), as well as between EMMPRIN and MMP-13 (r=0.474; p<0.001) and between EMMPRIN and IFN- (r=0.393; p=0.001). The higher quantity of myofibroblasts observed in solid ameloblastomas, odontogenic keratocysts and unicystic ameloblastomas suggests that these cells are one of the factors responsible for the more aggressive biological behavior of these tumors, although the myofibroblast population was not correlated with TGF-1, IFN-, MMP-13 or EMMPRIN. The correlation between MMP- 13 and TGF-1 suggests that the latter induces the expression of this metalloproteinase. The present results also support the well-established role of EMMPRIN as an inducer of MMP-13. Furthermore, the relationship between EMMPRIN and IFN- and between MMP-13 and IFN- suggests synergism in the antifibrotic effect of these markers

Elevated hyaluronan and extracellular matrix metalloproteinase inducer levels in women with preeclampsia

Purpose: Preeclampsia (PE) is a specific syndrome of pregnancy clinically identified by hypertension and proteinuria from the 20th week of gestation associated with a systemic inflammatory response and oxidative stress. While pro-inflammatory cytokines have been extensively studied in PE, other factors in the circulation that also influence the magnitude of inflammation have received much less attention. The present study compared serum concentrations of five immune-regulatory compounds in normotensive pregnant women and in women with gestational hypertension (GH) or PE. Methods: Sixty women with PE, 53 with GH and 40 normotensive women paired by gestational age were evaluated. Sera were evaluated for concentrations of extracellular matrix metalloproteinase inducer (EMMPRIN), hyaluronan, gelsolin, visfatin and histone 2B by ELISA. Differences between groups were analyzed by nonparametric tests, with a significance level of 5 %. Results: Increased levels of EMMPRIN and hyaluronan were present in preeclamptic women as compared to the GH and normotensive groups. There was no difference between groups in gelsolin, visfatin or histone 2B. Conclusion: Increased release of EMMPRIN and hyaluronan may contribute to an elevated pro-inflammatory response and tissue damage in women with PE. © 2013 Springer-Verlag Berlin Heidelberg.

Characterization of monocarboxylate transporters (MCTs) expression in soft tissue sarcomas: distinct prognostic impact of MCT1 sub-cellular localization

Background: Soft tissue sarcomas (STSs) are a group of neoplasms, which, despite current therapeutic advances, still confer a poor outcome to half of the patients. As other solid tumors, STSs exhibit high glucose consumption rates, associated with worse prognosis and therapeutic response. As highly glycolytic tumors, we hypothesized that sarcomas should present an increased expression of lactate transporters (MCTs).Methods: Immunohistochemical expression of MCT1, MCT2, MCT4 and CD147 was assessed in a series of 86 STSs and the expression profiles were associated with patients' clinical-pathological parameters.Results: MCT1, MCT4 and CD147 were mainly observed in the plasma membrane of cancer cells (around 60% for MCTs and 40% for CD147), while MCT2 was conspicuously found in the cytoplasm (94.2%). Importantly, we observed MCT1 nuclear expression (32.6%). MCT1 and MCT4, alone or co-expressed with CD147 in the plasma membrane, were associated with poor prognostic variables including high tumor grade, disease progression and shorter overall survival. Conversely, we found MCT1 nuclear expression to be associated with low grade tumors and longer overall survival.Conclusions: The present work represents the first report of MCTs characterization in STSs. We showed the original finding of MCT1 expression in the nucleus. Importantly, opposite biological roles should be behind the dual sub-cellular localization of MCT1, as plasma membrane expression of MCT1 is associated with worse patients' prognosis, while nuclear expression is associated with better prognosis.

Contribution of vascular resident mesenchymal stromal cells to abdominal aortic aneurysm pathogenesis: increased MMP-9 expression and ineffective immunomodulation

Background. Ageing and inflammation are critical for the occurrence of aortic diseases. Extensive inflammatory infiltrate and excessive ECM proteloysis, mediated by MMPs, are typical features of abdominal aortic aneurysm (AAA). Mesenchymal Stromal Cells (MSCs) have been detected within the vascular wall and represent attractive candidates for regenerative medicine, in virtue of mesodermal lineage differentiation and immunomodulatory activity. Meanwhile, many works have underlined an impaired MSC behaviour under pathological conditions. This study was aimed to define a potential role of vascular MSCs to AAA development. Methods. Aortic tissues were collected from AAA patients and healthy donors. Our analysis was organized on three levels: 1) histology of AAA wall; 2) detection of MSCs and evaluation of MMP-9 expression on AAA tissue; 3) MSC isolation from AAA wall and characterization for mesenchymal/stemness markers, MMP-2, MMP-9, TIMP-1, TIMP-2 and EMMPRIN. AAA-MSCs were tested for immunomodulation, when cultured together with activated peripheral blood mononuclear cells (PBMCs). In addition, a co-colture of both healthy and AAA MSCs was assessed and afterwards MMP-2/9 mRNA levels were analyzed. Results. AAA-MSCs showed basic mesenchymal properties: fibroblastic shape, MSC antigens, stemness genes. MMP-9 mRNA, protein and enzymatic activity were significantly increased in AAA-MSCs. Moreover, AAA-MSCs displayed a weak immunosuppressive activity, as shown by PBMC ongoing along cell cycle. MMP-9 was shown to be modulated at the transcriptional level through the direct contact as well as the paracrine action of healthy MSCs. Discussion. Vascular injury did not affect the MSC basic phenotype, but altered their function, a increased MMP-9 expression and ineffective immunmodulation. These data suggest that vascular MSCs can contribute to aortic disease. In this view, the study of key processes to restore MSC immunomodulation could be relevant to find a pharmacological approach for monitoring the aneurysm progression.