Extracellular Matrix Metalloproteinase Inducer is Regulated Developmentally and Functionally in the Rat Submandibular Gland

The salivary gland undergoes complex process of growth and differentiation of the branching morphogenesis of ductal system during the prenatal and early postnatal periods which are regulated by various elements in the extracellular matrix. Extracellular matrix metalloproteinase inducer (EMMPRIN) is a cell adhesion molecule. In the present study, localization and expression of EMMPRIN in development and effects of chorda-lingual denervation and cyclosporine A (CsA) treatment on the EMMPRIN expression were investigated. Immunohistochemistry, RT-PCR and Western blot were used to determine expression level. Immunohistochemistry revealed that EMMPRIN was localized specifically in the cytoplasm of ductal cells, not acini of the submandibular gland all the postnatal periods. At prenatal day 18, when the formation of ducts was not definite, no immunoreactivity was observed. Both Western blot and RT-PCR analyses revealed that EMMPRIN expression was maintained up to postnatal day 7, decreased after postnatal day 10. The EMMPRIN expression was upregulated by the surgical denervation of the chorda-lingual nerve in the gland as well as by the CsA treatment. The present study suggests that EMMPRIN is a crucial molecule for maintaining physiological functions of the salivary gland.

The expression and significance of Slug,EMMPRIN and E-cadherin in salivary adenoid cystic carcinoma / 实用口腔医学杂志

Objective:To investigate the expression of Slug,EMMPRIN and E-cadherin in salivary adenoid cystic carcinoma (SACC)and its correlation with clinicopathological characteristics,and the correlation among themselves.Methods:Slug,EMMPRIN and E-cadherin expression in 1 1 5 SACC cases of SACC was examined by immunohistochemical staining.The results and clinicopatho-logical data were statistically analyzed.Results:High positive expression frequencies of Slug(76.5%)and EMMPRIN(69.6%)and low positive expression frequency of E-cadherin(51 .3%)were found in 1 1 5 SACC cases.The expression of Slug and EMMPRIN was positively associated with the histopathological types,clinical stages,perineural invasion,recurrence and distance metastasis(P <0.05).The expression of E-cadherin was negatively associated with the histopathological types,clinical stages,perineural invasion and distance metastasis(P <0.05).There was a significant correlation between Slug and EMMPRIN expression(P <0.05),negative correlation between EMMPRIN and E-cadherin expression(P <0.05)and between Slug and E-cadherin expression(P <0.05).Con-clusion:The expression of Slug,EMMPRIN and E-cadherin is closely correlated to the clinicopathological characteristics of SACC.

Detecção de Periodontopatógenos, Glicoproteína Emmprin (Cd-147) e sua correlação com MMP-2 e MMP-9 / Detection of Periodontopathogens, Glycoprotein Emmprin (CD-147) and its Correlation whith MMP-2 and MMP-9

A periodontite é uma doença infecciosa caracterizada pela secreção de uma variedade de mediadores inflamatórios que levam a destruição dos tecidos de suporte dental e possível perda dos dentes, em associação com a infecção por múltiplas espécies bacterianas. Estima-se que mais de 400 espécies colonizam o biofilme dental e algumas das espécies bucais relacionadas à doença periodontal estejam no biofilme subgengival como Porphyromonas gingivalis, Tannerella forsythia, Treponema denticola. Entretanto, outros microrganismos podem estar relacionados a patologia desta doença, como Filifactor alocis e Prevotella tannerae. Esses microrganismos e seus subprodutos, como endotoxinas liberados o meio extracelular, levam ao estímulo da glicoproteína indutora de metaloproteinase (EMMPRIN, CD-147), que estimula a liberação de MMPs por fibroblastos e células endoteliais, levando a destruição do tecido. Com o objetivo de detectar F. alocis, P. tannerae e T. denticola, glicoproteina EMMPRIN (CD-147) e sua correlação com MMP-2 e MMP-9, amostras de fluido subgengival de pacientes com periodontite crônica, foram coletados de sítios sadios e doentes antes do tratamento periodontal básico e após 60 dias do tratamento. Os respectivos DNAs das bactérias foram extraídos e trechos do gene 16S foram amplificados e posteriormente realizados PCR convencional para a análise microbiológica dos microrganismos. Para a quantificação do EMMPRIN (CD-147), MMP-2 e MMP-9 foi usado ELISA-Sandwich. Resultados demonstraram que o grupo doente aumentou significantemente T. denticola, F. alocis e P. tannerae quando comparados com sítios saudáveis. MMP-2 e MMP-9 foram detectados em altas concentrações com redução estatisticamente significante após tratamento periodontal para MMP-2, mas não houve correlação com EMMPRIN.

Periodontitis is an infectious disease characterized by thesecretion of a variety of inflammatory mediators that lead todestruction of tooth supporting tissues, including the possibleloss of alveolar bone, in association with infection with multiplespecies of bacteria. It is estimated that more than 400 speciescolonize the biofilm and some oral species related to periodontaldisease is present in the subgingival including P. gingivalis, T.forsythia and T. denticola. However, other organisms may berelated of this disease, as Filifactor allocis and Prevotella tannerae.These microorganisms and subproducts such as endotoxinsreleased into the extracellular lead to the stimulation of metalloproteinaseinducer glycoprotein (EMMPRIN, CD-147), whichstimulates the release of MMPs by host cells, like fibroblasts andendothelial cells, thus leading to tissue destruction. The objectiveof this study was to detect F. allocis, P. tannerae, T. denticolaand the glycoprotein EMMPRIN (CD-147) and its correlationwith MMP-2 and MMP-9 in subgingival fluid samples of patientswith chronic periodontitis. Fluids were collected from healthyand disease subgingival sites of 20 healthy individuals beforebasic periodontal treatment and after of 60 days of treatment.Their DNAs were extracted and portions of the 16S gene wereamplified and performed conventional PCR. For immunologicalanalysis and quantification of EMMPRIN (CD-147), MMP-2 andMMP-9 was used ELISA-Sandwich. Results demonstrated thatthe 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 withstatistically significantly reduction after periodontal treatment toMMP-2, but without correlation with EMMPRIN.

Expression and significance of MMP-14 and EMMPRIN in human oral squamous cell carcinoma / 安徽医科大学学报

Objective To investigate the expression and significance of MMP-14 and EMMPRIN and the correlation between them in oral squamous cell carcinoma ( OSCC) . Methods The expression of MMP-14 and EMMPRIN mR-NA in 43 cases of OSCC and 11 cases of normal oral tissue were detected by RT-PCR. Results The expression of MMP-14 and EMMPRIN mRNA in OSCC was significantly higher than those in normal oral tissue ( P <0.05 ) . Compared with those without lymph node metastasis in OSCC, the expression of MMP-14 and EMMPRIN mRNA with lymph node metastasis was significantly higher ( P <0.05 ) . Furthermore, the expression of MMP-14 and EMMPRIN was significantly associated with pathological stage and the lymph node metastasis,but neither of them was correlated with sex and age. Correlation analysis showed that MMP-14 expression level was positively related to EMMPRIN level(r=0.801,P<0.01). Conclusion The expression of MMP-14 and EMMPRIN in OSCC is signif-icantly correlated with occurrence, progress and metastasis of OSCC. Therefore, the combination of detecting the expression of MMP-14 and EMMPRIN mRNA may evaluate the degree of malignancy and predict the prognosis of patients with OSCC.

Expressions of CD147 and MMP-9 in laryngeal squamous cell carcinoma and their significance / 西安交通大学学报(医学版)

Objective To investigate the expressions of CD147 and matrix metalloproteinases-9 (MMP-9)in laryngeal squamous cell carcinoma (LSCC)tissue and their clinical significance.Methods The expressions of CD147 and MMP-9 were analyzed semi-quantitatively by immunohistochemical staining in LSCC and control group tissues.Results ① The positive rate of CD147 was 83.3% (30/36)in LSCC,which was higher than that in laryngeal polyp (33.3%,5/15)and in adjacent normal tissue (16.7%,6/36);it was related to histological grade, clinical stage and lymph node metastasis status (P<0 .0 5 ).② The positive rate of MMP-9 was 7 2 .2% (2 6/3 6 )in LSCC,which was higher than that in laryngeal polyp (13.3%,2/15)and in adjacent normal tissue (5.6%,2/36);it was related to histological grade,T stage,clinical stage and lymph node metastasis status (P<0.05).③ There was a positive correlation between the expressions of CD147 and MMP-9 in LSCC tissue (r=0.721,P=0.000). Conclusion The over-expressions of CD147 and MMP-9 in LSCC may contribute to the development and metastasis of LSCC.

Estudo imuno-histoquímico da presença de miofibroblastos e da expressão do fator transformador de crescimento-beta1, interferon gama, mataloproteinase de matriz 13 e indutor de metaloproteinases de matriz em lesões odontogênicas epiteliais / Immunohistochemical study of the presence of myofibroblasts and the expression of transforming growth factor-beta1, interferon gamma, mataloproteinase matrix 13 and matrix metalloproteinase inducer in odontogenic epithelial lesions

Os miofibroblastos são células que apresentam um fenótipo híbrido exibindo características morfológicas de fibroblastos e de células musculares lisas, sendo a aquisição de tal fenótipo denominada diferenciação, passando então a expressar a a-SMA, a qual é importante na identificação dessas células. Estudos têm sugerido que os miofibrobíastos apresentam relação com a agressividade de diversas lesões e que o seu processo de diferenciação estaria relacionado à expressão do TGF-pl e do IFN-y atuando, respectivamente, no estímulo e na inibição dessa diferenciação. O objetivo deste trabalho foi investigar o papel dos miofibroblastos em lesões odontogênicas epiteliais, relacionando-os à agressividade das lesões e analisar por meio da imuno-histoquímica. a expressão do TGF-pl e IFN-y no processo de diferenciação, além da análise da MMP-13 que é ativada por miofibroblastos e do indutor de metaloproteinases de matriz (EMMPRIN) como precursor desta MMP. A amostra foi constituída por 20 ameloblastomas sólidos, 10 ameloblastomas unicfsticos, 20 ceratocistos odontogênicos e 20 tumores odontogênícos adenomatóides. Para a avaliação dos miofibroblastos, foram quantificadas as células imunorreativas ao anticorpo a-SMA presentes no tecido conjuntivo, próximo ao tecido epitelial. As expressões de TGF-pl, IFN-y, MMP-13 e EMMPRIN, foram avaliadas no componente epitelial e no conjuntivo, estabelecendo-se o percentual de imunorreatividade e atribuindo-se escores de 0 a 4. A análise dos miofibroblastos evidenciou maior concentração nos ameloblastomas sólidos (média de 30,55), seguido pelos ceratocistos odontogênicos (22,50), ameloblastomas unicísticos (20,80) e tumores odontogênicos adenomatóides (19,15) com valor de p= 0,001. Não foi encontrada correlação significativa entre TGF-pl e IFN-y no processo de diferenciação dos miofibroblastos, bem como na relação entre a quantidade de miofibroblastos e a expressão da MMP-13. Constatou-se, correlação estatística entre MMP-13 e TGF-pi (r= 0,087; p= 0,011) além de significante correlação entre MMP-13 e IFN-y (r=0,348; p=0,003). Entre EMMPRÍN e MMP-13 verificou-se significância (r= 0,474; p<0,001) assim como entre EMMPRIN e IFN-y (r=0,393; p=0,001). A maior quantidade de miofibroblastos evidenciada nos ameloblastomas sólidos, ceratocistos odontogênicos e ameloblastomas unicísticos sugere que estas células podem ser um dos fatores responsáveis para um comportamento biológico mais agressivo destas lesões, embora a população de miofibroblastos não tenha apresentado correlação com TGF- -pi, IFN-y ,MMP-13 e EMMPRIN. Quanto a correlação evidenciada entre MMP-13 e TGF-pl, isto pode sugerir um papel indutor do TGF-pl para a expressão da MMP-13, assim como os resultados deste estudo reforçam a relação bem estabelecida do EMMPRIN como indutor da MMP-13. Constatou-se também relação entre EMMPRIN e IFN-y assim como entre MMP-13 e IFN-y sugerindo, dessa forma, um sinergismo na ação anti-fibrótica desses marcadores.

Myofibroblasts are cells that exhibit a hybrid phenotype, sharing the morphoíogical characteristics of fibroblasts and smooth muscle cells, which is acquired during a process called differentiation. These cells then start to express a-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-pl and IFN-y 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-pl and IFN-y 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-a-SMA-immunoreactive cells were quantified in connective tissue close to the epithelium. Immunoexpression of TGF-pl, IFN-y, MMP-13 and EMMPRIN was evaluaíed 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=0.00). No significant correlation between TGF-pl and IFN-y 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-pi (r=0.087; p=0.01 1), between MMP-13 and ÍFN-y (r=0.348; p=0.003), as well as between EMMPRIN and MMP-13 (r=0.474; /xO.001) and between EMMPRIN and IFN-y (r=0.393; p=0.00). 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-01, IFN-y, MMP-13 or EMMPRIN. The correlation between MMP-13 and TGF-pl 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-y and between MMP-13 and IFN-y suggests synergism in the antifibrotic effect of these markers.

Increased Expression of EMMPRIN and VEGF in the Rat Brain after Gamma Irradiation

The extracellular matrix metalloproteinase inducer (EMMPRIN) has been known to play a key regulatory role in pathological angiogenesis. A elevated activation of vascular endothelial growth factor (VEGF) following radiation injury has been shown to mediate blood-brain barrier (BBB) breakdown. However, the roles of EMMPRIN and VEGF in radiation-induced brain injury after gamma knife surgery (GKS) are not clearly understood. In this study, we investigated EMMPRIN changes in a rat model of radiation injury following GKS and examined potential associations between EMMPRIN and VEGF expression. Adult male rats were subjected to cerebral radiation injury by GKS under anesthesia. We found that EMMPRIN and VEGF expression were markedly upregulated in the target area at 8-12 weeks after GKS compared with the control group by western blot, immunohistochemistry, and RT-PCR analysis. Immunofluorescent double staining demonstrated that EMMPRIN signals colocalized with caspase-3 and VEGF-positive cells. Our data also demonstrated that increased EMMPRIN expression was correlated with increased VEGF levels in a temporal manner. This is the first study to show that EMMPRIN and VEGF may play a role in radiation injuries of the central nervous system after GKS.

Expression of EMMPRIN, MMPs and TIMP2 in retinoblastoma and normal retinal tissue / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the expression of EMMPRIN, MMP1, MMP9 and TIMP2 in retinoblastoma (RB) and normal retinal tissues and their clinicopathological significance and interrelationship.METHODS: Envision immunohistochemistry stainings of EMMPRIN, MMP1, MMP9 and TIMP2 were performed in 30 enucleated eyeballs with retinoblastoma and 15 specimens of normal retina tissue, which had been routinely imbedded with paraffin.RESULTS: Positive rate of EMMPRIN, MMP1, MMP9 expression was higher in RB tissue than in normal control (P<0.01), while TIMP2 expression was lower in RB than in normal retinal tissue (P<0.01). Samples from RB cases of clinical stage Ⅰ, differentiated type, and life span≥2 years had lower positive rate in expression of EMMPRIN, MMP1, MMP9 than those from RB cases of clinical stage Ⅲ, undifferentiated type, and life span<2 years (P<0.05 or P<0.01), while samples from RB cases of differentiated type, optic nerve unaffected, and life span≥2 years had markedly higher positive rate in expression of TIMP2 than those from RB cases of undifferentiated type, optic nerve involved and life span<2 years (P<0.05 or P<0.01). In RB tissues, EMMPRIN, MMP1, MMP9 expressions were highly consistent (P<0.05), whereas TIMP2 expression is highly inconsistent with EMMPRIN, MMP1, MMP9 expression levels (P<0.05).CONCLUSION: The expression level of EMMPRIN, MMP1, MMP9 and TIMP2 may be an important marker of RB progression, invasion and prognosis. There exist internally mutual regulation relations among them.

Expression of MT1-MMP, EMMPRIN, P-gp in gastric carcinoma and their correlation / 中国癌症杂志

Purpose:To study the expression of MT1-MMP ,EMMPRIN and P-gp in gastric carcinoma and the relation of invasion, metastasis with drug resistance in gastric carcinoma. Methods:Detected membrane-type-1 matrix metalloproteinase (MT1-MMP), extra-cellular matrix metalloproteinase inducer (EMMPRIN), P-glycoprotein (P-gp) in gastric carcinoma by immunohistochemical method. Results:Among 40 cases of gastric carcinoma, there were 15 cases (37.5%) MT1-MMP-positive, 26 cases (65%) EMMPRIN-positive, 23 cases (57.5%) P-gp-positive respectively. The over-expression of MT1-MMP, EMMPRIN, P-gp was associated with invasive depth and lymph node matasteses of tumor cells(P