Tofacitinib Regulates Endostatin via Effects on CD147 and Cathepsin S.

Angiogenesis is critical for rheumatoid arthritis (RA) progression. The effects of tofacitinib, a JAK-STAT inhibitor used for RA treatment, on angiogenesis in RA are unclear. We, therefore, evaluated the levels of angiogenic factors in two systems of a human co-culture of fibroblast (HT1080) and monocytic (U937) cell lines treated with tofacitinib and in serum samples from RA patients before and after six months of tofacitinib treatment. Tofacitinib reduced CD147 levels, matrix metalloproteinase-9 (MMP-9) activity, and angiogenic potential but increased endostatin levels and secreted proteasome 20S activity. In vitro, tofacitinib did not change CD147 mRNA but increased miR-146a-5p expression and reduced STAT3 phosphorylation. We recently showed that CD147 regulates the ability of MMP-9 and secreted proteasome 20S to cleave collagen XVIIIA into endostatin. We show here that tofacitinib-enhanced endostatin levels are mediated by CD147, as CD147-siRNA or an anti-CD147 antibody blocked proteasome 20S activity. The correlation between CD147 and different disease severity scores supported this role. Lastly, tofacitinib reduced endostatin' s degradation by inhibiting cathepsin S activity and recombinant cathepsin S reversed this in both systems. Thus, tofacitinib inhibits angiogenesis by reducing pro-angiogenic factors and enhancing the anti-angiogenic factor endostatin in a dual effect mediated partly through CD147 and partly through cathepsin S.

The Roles of GDF-9, BMP-15, BMP-4 and EMMPRIN in Folliculogenesis and In Vitro Fertilization.

Growth differentiation factor 9 (GDF-9) contributes to early ovarian development and oocyte survival. Higher concentrations of GDF-9 in follicular fluid (FF) are associated with oocyte nuclear maturation and optimal embryo development. In in vitro fertilization (IVF), GDF-9 affects the ability of the oocyte to fertilize and subsequent embryonic development. Bone morphogenetic protein 15 (BMP-15) is involved in the regulation of ovarian function and affects oocyte development. During IVF, BMP-15 contributes to the formation of competent blastocysts. BMP-15 may play a role in embryo implantation by affecting endometrial receptivity. Bone morphogenetic protein 4 (BMP-4) is involved in the regulation of follicle growth and development and affects granulosa cell (GC) differentiation. In relation to IVF, BMP-4 is important for embryonic development, influences cell fate and differentiation, and plays a role in facilitating embryo-endometrial interactions during the implantation process. Extracellular matrix metalloproteinase inducer (EMMPRIN) is associated with ovulation and follicle rupture, promotes the release of mature eggs, and affects the modification of the extracellular matrix of the follicular environment. In IVF, EMMPRIN is involved in embryo implantation by modulating the adhesive properties of endometrial cells and promotes trophoblastic invasion, which is essential for pregnancy to occur. The purpose of the current article is to review the studies and recent findings of GDF-9, BMP-15, BMP-4 and EMMPRIN as fundamental factors in normal follicular development and in vitro fertilization.

Pseudolaric Acid B Targets CD147 to Selectively Kill Acute Myeloid Leukemia Cells.

Acute myeloid leukemia (AML) is an aggressive blood cancer. With low survival rates, new drug targets are needed to improve treatment regimens and patient outcomes. Pseudolaric acid B (PAB) is a plant-derived bioactive compound predicted to interact with cluster of differentiation 147 (CD147/BSG). CD147 is a transmembrane glycoprotein overexpressed in various malignancies with suggested roles in regulating cancer cell survival, proliferation, invasion, and apoptosis. However, the detailed function of PAB in AML remains unknown. In this study, AML cell lines and patient-derived cells were used to show that PAB selectively targeted AML (IC50: 1.59 ± 0.47 µM). Moreover, proliferation assays, flow cytometry, and immunoblotting confirmed that PAB targeting of CD147 resulted in AML cell apoptosis. Indeed, the genetic silencing of CD147 significantly suppressed AML cell growth and attenuated PAB activity. Overall, PAB imparts anti-AML activity through transmembrane glycoprotein CD147.

EMMPRIN promotes spheroid organization and metastatic formation: comparison between monolayers and spheroids of CT26 colon carcinoma cells.

Background: In vitro studies often use two-dimensional (2D) monolayers, but 3D cell organization, such as in spheroids, better mimics the complexity of solid tumors. To metastasize, cancer cells undergo the process of epithelial-to-mesenchymal transition (EMT) to become more invasive and pro-angiogenic, with expression of both epithelial and mesenchymal markers. Aims: We asked whether EMMPRIN/CD147 contributes to the formation of the 3D spheroid structure, and whether spheroids, which are often used to study proliferation and drug resistance, could better model the EMT process and the metastatic properties of cells, and improve our understanding of the role of EMMPRIN in them. Methods: We used the parental mouse CT26 colon carcinoma (CT26-WT) cells, and infected them with a lentivirus vector to knock down EMMPRIN expression (CT26-KD cells), or with an empty lentivirus vector (CT26-NC) that served as a negative control. In some cases, we repeated the experiments with the 4T1 or LLC cell lines. We compared the magnitude of change between CT26-KD and CT26-WT/NC cells in different metastatic properties in cells seeded as monolayers or as spheroids formed by the scaffold-free liquid overlay method. Results: We show that reduced EMMPRIN expression changed the morphology of cells and their spatial organization in both 2D and 3D models. The 3D models more clearly demonstrated how reduced EMMPRIN expression inhibited proliferation and the angiogenic potential, while it enhanced drug resistance, invasiveness, and EMT status, and moreover it enhanced cell dormancy and prevented CT26-KD cells from forming metastatic-like lesions when seeded on basement membrane extract (BME). Most interestingly, this approach enabled us to identify that EMMPRIN and miR-146a-5p form a negative feedback loop, thus identifying a key mechanism for EMMPRIN activities. These results underline EMMPRIN role as a gatekeeper that prevents dormancy, and suggest that EMMPRIN links EMT characteristics to the process of spheroid formation. Conclusions: Thus, 3D models can help identify mechanisms by which EMMPRIN facilitates tumor and metastasis progression, which might render EMMPRIN as a promising target for anti-metastatic tumor therapy.

Glucose Promotes EMMPRIN/CD147 and the Secretion of Pro-Angiogenic Factors in a Co-Culture System of Endothelial Cells and Monocytes.

Vascular complications in Type 2 diabetes mellitus (T2DM) patients increase morbidity and mortality. In T2DM, angiogenesis is impaired and can be enhanced or reduced in different tissues ("angiogenic paradox"). The present study aimed to delineate differences between macrovascular and microvascular endothelial cells that might explain this paradox. In a monoculture system of human macrovascular (EaHy926) or microvascular (HMEC-1) endothelial cell lines and a monocytic cell line (U937), high glucose concentrations (25 mmole/L) increased the secretion of the pro-angiogenic factors CD147/EMMPRIN, VEGF, and MMP-9 from both endothelial cells, but not from monocytes. Co-cultures of EaHy926/HMEC-1 with U937 enhanced EMMPRIN and MMP-9 secretion, even in low glucose concentrations (5.5 mmole/L), while in high glucose HMEC-1 co-cultures enhanced all three factors. EMMPRIN mediated these effects, as the addition of anti-EMMPRIN antibody decreased VEGF and MMP-9 secretion, and inhibited the angiogenic potential assessed through the wound assay. Thus, the minor differences between the macrovascular and microvascular endothelial cells cannot explain the angiogenic paradox. Metformin, a widely used drug for the treatment of T2DM, inhibited EMMPRIN, VEGF, and MMP-9 secretion in high glucose concentration, and the AMPK inhibitor dorsomorphin enhanced it. Thus, AMPK regulates EMMPRIN, a key factor in diabetic angiogenesis, suggesting that targeting EMMPRIN may help in the treatment of diabetic vascular complications.

Paradoxical Changes: EMMPRIN Tissue and Plasma Levels in Marfan Syndrome-Related Thoracic Aortic Aneurysms.

Background: Thoracic aortic aneurysms (TAAs) associated with Marfan syndrome (MFS) are unique in that extracellular matrix metalloproteinase inducer (EMMPRIN) levels do not behave the way they do in other cardiovascular pathologies. EMMPRIN is shed into the circulation through the secretion of extracellular vesicles. This has been demonstrated to be dependent upon the Membrane Type-1 MMP (MT1-MMP). We investigated this relationship in MFS TAA tissue and plasma to discern why unique profiles may exist. Methods: Protein targets were measured in aortic tissue and plasma from MFS patients with TAAs and were compared to healthy controls. The abundance and location of MT1-MMP was modified in aortic fibroblasts and secreted EMMPRIN was measured in conditioned culture media. Results: EMMPRIN levels were elevated in MFS TAA tissue but reduced in plasma, compared to the controls. Tissue EMMPRIN elevation did not induce MMP-3, MMP-8, or TIMP-1 expression, while MT1-MMP and TIMP-2 were elevated. MMP-2 and MMP-9 were reduced in TAA tissue but increased in plasma. In aortic fibroblasts, EMMPRIN secretion required the internalization of MT1-MMP. Conclusions: In MFS, impaired EMMPRIN secretion likely contributes to higher tissue levels, influenced by MT1-MMP cellular localization. Low EMMPRIN levels, in conjunction with other MMP analytes, distinguished MFS TAAs from controls, suggesting diagnostic potential.

Soluble CD147 regulates endostatin via its effects on the activities of MMP-9 and secreted proteasome 20S.

During progression of rheumatoid arthritis (RA), angiogenesis provides oxygen and nutrients for the cells' increased metabolic demands and number. To turn on angiogenesis, pro-angiogenic factors must outweigh anti-angiogenic factors. We have previously shown that CD147/extracellular matrix metalloproteinase inducer (EMMPRIN) can induce the expression of the pro-angiogenic factors vascular endothelial growth factor (VEGF) and matrix metallopeptidase 9 (MMP-9) in a co-culture of the human HT1080 fibrosarcoma and U937 monocytic-like cell lines. However, whether CD147 influences anti-angiogenic factors was not known. We now show that relative to single cultures, the co-culture of these cells not only enhanced pro-angiogenic factors but also decreased the anti-angiogenic factors endostatin and thrombospondin-1 (Tsp-1), generally increasing the angiogenic potential as measured by a wound assay. Using anti-CD147 antibody, CD147 small interfering RNA (siRNA), and recombinant CD147, we demonstrate that CD147 hormetically regulates the generation of endostatin but has no effect on Tsp-1. Since endostatin is cleaved from collagen XVIII (Col18A), we applied different protease inhibitors and established that MMP-9 and proteasome 20S, but not cathepsins, are responsible for endostatin generation. MMP-9 and proteasome 20S collaborate to synergistically enhance endostatin generation, and in a non-cellular system, CD147 enhanced MMP-9 activity and hormetically regulated proteasome 20S activity. Serum samples obtained from RA patients and healthy controls mostly corroborated these findings, indicating clinical relevance. Cumulatively, these findings suggest that secreted CD147 mediates a possibly allosteric effect on MMP-9 and proteasome 20S activities and can serve as a switch that turns angiogenesis on or off, depending on its ambient concentrations in the microenvironment.

Targeting cancer-derived extracellular vesicles by combining CD147 inhibition with tissue factor pathway inhibitor for the management of urothelial cancer cells.

BACKGROUND: Extracellular vesicles (EVs), including microvesicles, hold promise for the management of bladder urothelial carcinoma (BLCA), particularly because of their utility in identifying therapeutic targets and their diagnostic potential using easily accessible urine samples. Among the transmembrane glycoproteins highly enriched in cancer-derived EVs, tissue factor (TF) and CD147 have been implicated in promoting tumor progression. In this in vitro study, we explored a novel approach to impede cancer cell migration and metastasis by simultaneously targeting these molecules on urothelial cancer-derived EVs. METHODS: Cell culture supernatants from invasive and non-invasive bladder cancer cell lines and urine samples from patients with BLCA were collected. Large, microvesicle-like EVs were isolated using sequential centrifugation and characterized by electron microscopy, nanoparticle tracking analysis, and flow cytometry. The impact of urinary or cell supernatant-derived EVs on cellular phenotypes was evaluated using cell-based assays following combined treatment with a specific CD147 inhibitor alone or in combination with a tissue factor pathway inhibitor (TFPI), an endogenous anticoagulant protein that can be released by low-molecular-weight heparins. RESULTS: We observed that EVs obtained from the urine samples of patients with muscle-invasive BLCA and from the aggressive bladder cancer cell line J82 exhibited higher TF activity and CD147 expression levels than did their non-invasive counterparts. The shedding of GFP-tagged CD147 into isolated vesicles demonstrated that the vesicles originated from plasma cell membranes. EVs originating from invasive cancer cells were found to trigger migration, secretion of matrix metalloproteinases (MMPs), and invasion. The same induction of MMP activity was replicated using EVs obtained from urine samples of patients with invasive BLCA. EVs derived from cancer cell clones overexpressing TF and CD147 were produced in higher quantities and exhibited a higher invasive potential than those from control cancer cells. TFPI interfered with the effect when used in conjunction with the CD147 inhibitor, further suppressing homotypic EV-induced migration, MMP production, and invasion. CONCLUSIONS: Our findings suggest that combining a CD147 inhibitor with low molecular weight heparins to induce TFPI release may be a promising therapeutic approach for urothelial cancer management. This combination can potentially suppress the tumor-promoting actions of cancer-derived microvesicle-like EVs, including collective matrix invasion.

Small particles or vesicles released by cancer cells into their surroundings have the potential to stimulate the spread and growth of cancer cells. In this study, we focused on two specific molecules presented by these cancer cell-derived vesicles that could play a role in promoting the dissemination of cancer cells: a protein related to blood clotting and a protein on the cell surface.We found that large vesicles from bladder cancer cells that have the ability to spread had higher levels of these proteins than vesicles from nonspreading cancer cells. We also found that the former could make cancer cells move about more, produce more of a substance that helps cancer cells spread, and invade other tissues.To counteract the cancer-promoting actions of these vesicles, we examined the impact of combining a naturally occurring anticlotting protein that can be released by medications derived from heparin with an inhibitor targeting the cancer cell surface protein. We found that this combination stopped the vesicles from helping cancer cells move about more, produce more of the spreading substance, and invade other tissues.This approach of simultaneously targeting the two protein molecules present on cancer cell-derived vesicles might be a new way to treat bladder cancer.

A Single-Center Retrospective Analysis of Kaposi's Sarcoma: Is There a Relationship Between Emmprin/CD147 Expression and Biological Behavior?

OBJECTIVES: Emmprin (CD147/BSG) protein is estimated to play a key role in cell migration and chemoresistance in viral carcinogenesis. However, there are very limited studies investigating the CD147 in the oncogenesis of Kaposi's sarcoma-associated herpesvirus. This study aims to reveal the relationship between CD147 expression with histopathological parameters, disease pattern, and recurrence in Kaposi's sarcoma (KS). METHODS: The study included 67 patients diagnosed with KS between January 1982 and September 2023. Clinical and histopathological features were analyzed retrospectively. HHV-8, CD31, and CD147 expressions were evaluated by immunohistochemistry. RESULTS: Sixteen (24%) female and 51 (76%) male patients with median age of 64 (10-86) were included in the study. CD147 was positive in 57 (85%) cases and associated with nodular pattern (P = .001), presence of solid/fibrosarcomatous area (P = .005), and high mitotic activity (P = .035). The disease relapsed in 17 (27%) of the 63 patients with median 2 (0-12) years follow-up. While a 5-year relapse-free survival was 48.5% in the CD147 diffuse positive group, it was 83.4% in focal positive and 100% in negative cases (P = .029). CONCLUSION: Our study exhibited the relationship between CD147 overexpression and recurrence in KS, but the inhomogeneity of the treatment groups and the small number of patients should also be considered. These findings may provide insight into the pathogenesis of KS and the development of targeted therapies in the future.

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.

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.

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.

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