Diagnostic value of combined detection of MT1-MMP, LncRNA SNHG15 and YAP in thyroid carcinoma / 国际检验医学杂志

Objective To explore the diagnostic value of combined detection of MT1-MMP, LncRNA SNHG15and YAP in thyroid carcinoma.Methods A total of 186thyroid carcinoma patients from July 2015to July 2017were selected as the observation group and 50healthy patients were selected as control group.The expression levels of MT1-MMP, LncRNA SNHG15, and YAP in tissues of the two groups were compared.The relationships between the clinical pathological parameters of thyroid carcinoma and the expression levels of MT1-MMP, LncRNA SNHG15, YAP were analyzed.Results The positive rate of MT1-MMP, YAP and LncRNA SNHG15in observation group were significantly higher than those in control group (P<0.05) .The expression level of YAP in tumors larger than 2cm diameter was significantly higher than that in tumors less than 2cm (P<0.05) .The expression level of YAP and LncRNA SNHG15with envelope infiltration were significantly higher than those without envelope infiltration (P<0.05) .The expression level of MT1-MMP in lymph node metastasis group was significantly higher than that in non-lymphatic metastasis group (P<0.05) .The expression level of YAP in gradesⅢandⅣof TNM were significantly higher than those in gradesⅠandⅡ (P<0.05) .Conclusion High expression level of MT1-MMP is associated with infiltration of thyroid carcinoma capsules and lymphatic metastasis.YAP is associated with greater proliferative capacity of papillary thyroid carcinoma, and LncRNA SNHG15is closely associated with infiltration of capsules.

Changes of lymphatic vessel density in lung adenocarcinoma in situ, minimally invasive adenocarcinoma, and invasive adenocarcinoma and the regulatory factors / 南方医科大学学报

OBJECTIVE@#To analyze the changes in tumor lymphatic vessel density (LVD) in patients with lung adenocarcinoma in situ (AIS), minimally invasive adenocarcinoma (MIA), and invasive adenocarcinoma (IA) and explore the regulatory factors of LVD.@*METHODS@#Complete clinicopathological data were collected form a total of 301 patients with lung adenocarcinoma, including 28 (9.3%) with AIS, 86 (28.6%) with MIA, and 187 (62.1%) with IA. The LVD of all the adenocarcinomas were calculated after D2-40 immunohistochemical staining, and MT1-MMP and VEGF-C expression levels were also evaluated. The differences in LVD among the groups and the correlations of tumor LVD with the expressions of MT1-MMP and VEGF-C and the clinicopathological factors were analyzed.@*RESULTS@#The LVD differed significantly among AIS, MIA, and IA groups (= 0.000). The LVDs was significantly correlated with the level of VEGF-C protein expression (=0.917, =0.009), tumor size (= 0.686, =0.017), lymph node metastasis (=0.739, =0.000), and clinical stage (=0.874, =0.012) of the patients.@*CONCLUSIONS@#Tumor lymphangiogenesis plays an important role in lung adenocarcinoma progression, and VEGF-C may promote this process.

Inhibition effect of curcumin on proliferation and invasion of breast cancer cell by decreasing expression of MT1-MMP / 实用医学杂志

Objective To investigate whether MT1-MMP is involved in the inhibition effect of curcumin on the proliferation and invasion of breast cancer cell and the mechanism . Methods Firstly, MCF-7 cell lines transfected by MT1-MMP eukaryotic expression vector was established. We divided all cells into 3 groups,including null vector transfection group, non-transfected and transfected group with different concentrations of curcumin. The expression of MT1-MMP protein, the proliferation and invasion ability were respectively analyzed by western blot, transwell method, and cell counting kit-8 (CCK-8). Results The expression of MT1-MMP was inhibited by curcumin. Transwell and CCK-8 experiment indicated the proliferation and invasion abilities of MT1-MMP transfected MCF-7 cells were inhibited by curcumin in a concentration dependent manner. Conclusion The inhibition value of curcumin on proliferation and invasion is probably due to its ability to inhibit the expression of MT1-MMP.

Expression of CD151 and MT1-MMP in adenocarcinoma of esophagogastric junction and their significances / 临床与实验病理学杂志

Purpose To investigate the expression of the protein of CD151 and MT1-MMP in adenocarcinoma of esophagogastric junc-tion tissues and to explore their relationship with the invasiveness and metastasis of the tumor. Methods CD151 and MT1-MMP pro-tein were detected by immunohistochemical staining respectively in 15 cases of paraneoplastic normal gastric mucosa tissue, 40 cases dysplasia and 172 cases of adenocarcinoma of esophagogastric junction tissues. Results All of the expression level of CD151 and MT1-MMP protein were increasing according to the order of normal-dysplsia-carcinoma. The expression rate of CD151 was 6. 67%, 20. 0% and 78. 3%, while the rate of MT1-MMP with 13. 3%, 22. 5% and 79. 1% in the normal, dysplasia and carcinoma subgroup. The expression rate of each protein were significant difference among the three goroups (P<0. 05). In the adenocarcinoma of esopha-gogastric junction tissues, the expression of them in the subgroup of poorly-differentiated with serosa invasion and lymph nodes metasta-sis was significantly higher than the other subgroup of well-differentiated with non serosa invasion or lymph nodes metastasis ( P <0. 05 ) . There was a positive correlation between the expression of CD151 and MT1-MMP protein in adenocarcinoma of esophagogastric junction tissues ( P<0. 05 ) . Conclusions CD151 and MT1-MMP assuredly and highly express in the adenocarcinoma of esopha-gogastric junction tissues and they have close relationships, which could involved in the invasiveness and metastasis synergistically in this tumor.

The Effects of Recombinant Synucleins and Insulin-like Growth Factor 1 on Cancer Cell Migration

The synuclein family consists of three distinct genes, alpha-synuclein, beta-synuclein, and gamma-synuclein. The alpha-synuclein and beta-synuclein are predominately expressed in brain and especially alpha-synuclein is related with Parkinson's disease, Alzheimer's disease, and dementia with Lewy bodies. The gamma-synuclein was first identified as breast cancer specific gene 1. It is expressed in the peripheral nervous system and also detected in breast and ovarian cancers. The gamma-synuclein is also known to mediate metastasis of breast and ovarian cancer cells. Insulin-like growth factor 1 (IGF-I) is one of the growth factors that plays an important role in cell proliferation and migration in cancer cells, as well as in normal cells. In this study, we investigated the migrations of SKOV-3, MDAMB-231, and HeLa cells by the recombinant synuclein proteins (alpha-, beta-, and gamma-synucleins) and IGF-I and the molecular mechanism. Furthermore, we investigated the membrane ruffle formation of SKOV-3 cells by recombinant synuclein proteins and IGF-I. As a result, synucleins and IGF-I were found to induce cancer cell migrations. Simultaneous synucleins and IGF-I treatment on the cancer cells induced more migrations than the individual synuclein or IGF-I treatments. The synucleins or IGF-I treatments increased the expressions of membrane-type1 matrix metalloproteinase (MT1-MMP) and cluster of differentiation 44 (CD44). Moreover, simultaneous synucleins and IGF-I treatments further increased the expressions of MT1-MMP and CD44. The synucleins and IGF-I promoted the conformational change of actin filaments, and then this led to the membrane ruffle formation.

Participación de MT1-MMP en la remodelación del ligamento periodontal durante la movilización dentaria / Role of MT1-MMP in the remodeling of the periodontal ligament during tooth movement

La movilización dentaria involucra una serie de cambios en los tejidos de soporte caracterizados por la activa remodelación de estos. La MT1-MMP o MMP-14 es una potente enzima proteolítica capaz de degradar colágeno tipo I, la principal molécula estructural del ligamento periodontal. La migración dentaria requiere de la degradación controlada del colágeno constituyente del ligamento periodontal. Sin embargo, no existen evidencias de la participación de MT1-MMP en la remodelación del tejido periodontal durante este proceso. En el presente estudio hemos analizado la expresión de MT1 -MMP y del marcador de actividad osteoclástica Fosfatasa Acida Tartrato Resistente (TRAP) en un modelo de migración dentaria en ratas. La migración dentaria fue activada mediante la inserción de una banda separadora entre los incisivos superiores. La expresión y distribución de TRAP y MT1-MMP fue evaluada a través de citoquímica e inmunohistoquímica a los días 1, 3, 5 y 7. La producción de TRAP fue identificada principalmente en osteoclastos ubicados en la zona de compresión del ligamento periodontal. La producción de MT1-MMP fue observada en fibroblastos de la zona de compresión del ligamento periodontal y osteoclastos ubicados en esta misma región. Nuestros resultados permiten proponer que tanto MT1 -MMP como TRAP participan en la remodelación de los tejidos de soporte periodontal durante la migración dentaria.

Tooth movement involves a series of changes of the supporting periodontal tissues characterized by the active connective tissue remodeling. MT1-MMP or MMP-14 belongs to the family of matrix metalloproteinases that are able to degrade type I collagen, the main molecule involved in periodontal attachment. Tooth migration requires the controlled degradation of periodontal ligament collagen fibers. However, evidences linking MT1 -MMP expression with periodontal tissue remodeling are lacking. In the present study, we have evaluated the expression of MT1-MMPand of the osteoclast marker Tartrate Resistant Acid Phosphatase (TRAP) in a model of tooth migration in rats. Tooth migration was induced after the insertion of a rubber band between the upper incisors. The distribution of TRAP and MT1 -MMP was evaluated by means of cytochemistry and immunohistochemistry respectively at days 1, 3, 5 and 7. TRAP production was identified in osteoclasts at the area of compression of the periodontal ligament. MT1-MMP distribution was observed in fibroblastsatthe compressed area of the periodontal ligament and also in osteoclasts of the same region. Our findings allow us to propose that MT1-MMP and TRAP take part of the tissue remodeling events observed during tooth movement.

The Role of PKCzeta on MT1-MMP Expression with Shear Stress and Cyclic Strain in Microvascular Endothelial Cells

PURPOSE: hear stress (SS) and cyclic strain (CS) influence the expression of membrane type 1-matrix metalloproteinase (MT1-MMP) in microvascular endothelial cells (MVECs). It is known that changes in the level of Sp1 phosphorylation are important for MT1-MMP expression following SS and CS. However, the exact mechanism underlying this process is poorly understood. The aim of this study was to determine the effect of PKCzeta on serine phosphorylation and activation of Sp1 in response to SS and CS. METHOD: MVECs were exposed to SS or CS for up to 8 hours with or without PKCzeta inhibitors. The activity and phosphorylation of Sp1 were assessed by Western blot analysis and immunoprecipitation. MT1-MMP protein expression was assessed by Western blot analysis. RESULT: PKCzeta was phosphorylated and activated under SS, whereas no significant changes were noted under CS. SS increased Sp1 phosphorylation in a time-dependent manner, but no changes in the Sp1 phosphorylation were observed when the MVECs were pretreated with the PKCzeta inhibitors. By contrast, MVECs exposed to CS in the presence or absence of PKCzeta inhibitors showed no change in the phosphorylation of Sp1. SS decreased MT1-MMP protein expression in a time-dependent manner, but in the presence of PKCzeta inhibitors, MT1-MMP expression was not changed compared with the static levels after SS. CS increases MT1-MMP expression in a time-dependent manner. Similar expression was observed when the cells were pretreated with PKCzeta inhibitors under CS. CONCLUSION: These data demonstrate that the increased affinity of Sp1 for the MT1-MMP's promoter site occurs because of PKCzeta induced phosphorylation of Sp1 in response to SS.

Egr-1 Mediates SiO2-driven Transcription of Membrane Type Ⅰ Matrix Metalloproteinase in Macrophages / 华中科技大学学报（医学）（英德文版）

The up-regulation mechanism of membrane type Ⅰ matrix metalloproteinase (MT1-MMP) in macrophages stimulated by silica in vitro and the contribution of early growth response 1 (Egr-1) transcription factor in the gene expression pathway were investigated. Macrophages stimulated by silica were treated with Egr-1 antibody or Egr-1 decoy oligodeoxynucleotides (ODN). The levels of MT1-MMP proteins were determined by Western blot and the expression of MT1-MMP mRNAs was detected by RT-PCR. The results showed as compared with control macrophages, silica-stimulated group showed up-regulated gene expression of MT1-MMP via Egr-1 (P＜0.01). Compared with silica-stimulated macrophages untreated with antibody, the cells treated with 5 μg/mL Egr-1 antibody were associated with reduced expression of MTI-MMP protein (P＜0.01) and mRNA (P＜0.01). Compared with silica-stimulated untransfected group, the Egr-1 "decoy" ODN group was associated with reduction in the expression of MT1-MMP protein and mRNA (P＜0.01). It was concluded gene expression of MT1-MMP which may play a critical role in silicosis was up-regulated by silica in macrophages. Egr-1 participated in the expression of MT1-MMP and positively regulated the expression. Both Egr-1 antibody and Egr-1 decoy ODN suppressed the expression of MT1-MMP through the Egr-1 pathway and may become a potential therapeutic tool in the management of silicosis in the future.

Expression of MT-1 MMP, MMP2, MMP9 and TIMP2 mRNAs in Ductal Carcinoma in Situ and Invasive Ductal Carcinoma of the Breast

We investigated the expression of membrane type-1 (MT1)-MMP, MMP2, MMP9 and TIMP2 mRNAs and their roles in ductal carcinoma in situ (DCIS) and T1 and T2 invasive ductal carcinoma of the breast. We further compared these two types of carcinomas for differences in microvessel density, and expression of angiogenic factors and CD44std. MT1-MMP, MMP2, MMP9 and TIMP2 mRNA were expressed in both DCIS and invasive ductal carcinomas. Expression rates of MT1-MMP, MMP2, MMP9 and TIMP2 mRNAs were not statistically different between DCIS and invasive ductal carcinomas, nor did they differ statistically when grouped by tumor size, histologic grade or nuclear grade of invasive ductal carcinoma. Microvessel density and expression of VEGF and TGF-beta were not statistically different between DCIS and invasive ductal carcinoma. CD44std expression was significantly increased in DCIS compared to invasive ductal carcinoma (p < 0.05) and it was also significantly increased in lower clinical stage, histologic grade and nuclear grade of invasive ductal carcinoma (p < 0.05). Axillary node metastasis was significantly correlated with MT1-MMP mRNA, VEGF and TGF-beta expression (p < 0.05) and MT1-MMP mRNA was positively correlated with VEGF expression and TIMP2 mRNA (p < 0.05). In summary, patterns of MMP mRNA expression in DCIS and invasive ductal carcinoma suggest that the invasive potential of breast carcinoma is already achieved before morphologically overt invasive growth is observed. As MT1-MMP mRNA expression is significantly correlated with axillary nodal metastasis, it may be useful as a prognostic indicator of invasive ductal carcinoma. Considering the positive correlation of MT1-MMP mRNA and TIMP2mRNA expression, our finding supports a role for TIMP2 in tumor growth, as well as the utility of CD44std as a prognostic indicator of breast cancer.

Expression of MT-1 MMP, MMP2, MMP9 and TIMP2 mRNAs in Ductal Carcinoma in Situ and Invasive Ductal Carcinoma of the Breast

We investigated the expression of membrane type-1 (MT1)-MMP, MMP2, MMP9 and TIMP2 mRNAs and their roles in ductal carcinoma in situ (DCIS) and T1 and T2 invasive ductal carcinoma of the breast. We further compared these two types of carcinomas for differences in microvessel density, and expression of angiogenic factors and CD44std. MT1-MMP, MMP2, MMP9 and TIMP2 mRNA were expressed in both DCIS and invasive ductal carcinomas. Expression rates of MT1-MMP, MMP2, MMP9 and TIMP2 mRNAs were not statistically different between DCIS and invasive ductal carcinomas, nor did they differ statistically when grouped by tumor size, histologic grade or nuclear grade of invasive ductal carcinoma. Microvessel density and expression of VEGF and TGF-beta were not statistically different between DCIS and invasive ductal carcinoma. CD44std expression was significantly increased in DCIS compared to invasive ductal carcinoma (p < 0.05) and it was also significantly increased in lower clinical stage, histologic grade and nuclear grade of invasive ductal carcinoma (p < 0.05). Axillary node metastasis was significantly correlated with MT1-MMP mRNA, VEGF and TGF-beta expression (p < 0.05) and MT1-MMP mRNA was positively correlated with VEGF expression and TIMP2 mRNA (p < 0.05). In summary, patterns of MMP mRNA expression in DCIS and invasive ductal carcinoma suggest that the invasive potential of breast carcinoma is already achieved before morphologically overt invasive growth is observed. As MT1-MMP mRNA expression is significantly correlated with axillary nodal metastasis, it may be useful as a prognostic indicator of invasive ductal carcinoma. Considering the positive correlation of MT1-MMP mRNA and TIMP2mRNA expression, our finding supports a role for TIMP2 in tumor growth, as well as the utility of CD44std as a prognostic indicator of breast cancer.

Construction of Antisense MT1-MMP Vector and Its Inhibitory Effects on Invasion of Human Ovarian Cancer Cells / 华中科技大学学报（医学）（英德文版）

Membrane-type 1 matrix metalloproteinase (MT1 MMP/MMP 14) plays crucial roles in tumor cell growth, invasion, and angiogenesis. To clarify whether the endogenously expressed MT1-MMP in metastatic human ovarian carcinoma cell lines SKOV3 plays a critical role in tumor cell invasiveness, antisense MT1-MMP cloned in eukaryotic expression vector pMMP14as was transferred into SKOV3 cells. 48h after transfection, decreased expression of endogenous MT1-MMP protein was detected in pMMP14as transfected SKOV3 cells and the activation of pro MMP2was inhibited markedly. The mean percentage of invasive cells was (62. 50 ±5. 30) % in pMMP14as-transfected cells, which was obviously less than that (97.20±6.90) % in the control.Thus, antisense MT1 MMP effectively inhibited the endogenous MT1 MMP expression and the invasiveness in SKOV3 cells, suggesting that MT1-MMP may be a therapeutic target molecule for human invasive ovarian cancers.

MT1-MMP and Egr-1 Expressions Induced by Different Strengths of Shear Stress in Endothelial Cell

PURPOSE: Membrane type-1-matrix metalloproteinase (MT1-MMP) plays a key role in endothelial cell (EC) migration, matrix remodeling, and angiogenesis. Previous studies demonstrated that a cyclic strain (CS) increases MT1-MMP expression by displacing specific protein 1(Sp1) with increased early growth response-1 (Egr-1) expression; and shear stress (SS) decreases MT1-MMP expression by Sp1 phosphorylation. However, the difference in MT1-MMP expression according to the change of SS is poorly understood. The aim of this study is to determine the effects of low or high SS on Egr-1 and MT1-MMP transcription and translation. METHOD: Bovine aortic ECs were exposed to oscillatory SS (low=0.1 dyne/cm2 or high=14 dyne/cm2) with orbital shaker for 0, 1, 4, or 8 hours. Activation of Egr-1 and MT1-MMP was assessed by the Northern blot and Western blot. RESULT: Although Egr-1 mRNA transcription and protein translation were induced (7.3-, 5.8-fold and 4.0-, 4.9-fold, respectively) in response to low SS (n=5, 0, 1, and 4 hr; P<0.05), MT1-MMP mRNA transcription and protein levels did not change remarkably. Egr-1 mRNA transcription and translation were induced (7.6-fold at 1 hr; 3.7- and 5.2-fold at 1 and 4 hr, respectively) in response to high SS (n=5; P<0.05). We observed that high SS decreased MT1-MMP mRNA transcription and translation in a time-dependent fashion (10%, 50%, and 90% reduction at 1, 4, and 8 hr, respectively; n=5, P<0.05). CONCLUSION: High SS induces Egr-1 up-regulation and inhibits MT1-MMP expression. But low SS has no effect on MT1-MMP expression in spite of Egr-1 up-regulation. These observations illustrate that the expression of MT1-MMP is dependent on, not only the type of hemodynamic forces, but also the strength of force (SS) in vascular endothelial cells.

Isolation and purification of protein complexes of MT1-MMP-hTAP fusion gene using tandem affinity purification system in chick fibroblast cells and evaluation of the expression effect / 第三军医大学学报

Objective To construct the fusion gene of membrane type matrix metalloproteinase1 humanized tandem affinity purification (MT1 MMP hTAP) and to evaluate the expression of its protein complexes isolated and purified using tandem affinity purification (TAP) system. Methods The MT1 MMP hTAP protein complexes were isolated and purified using the IgG agarose beads and calmodulin sepharose beads affinity chromatography according to the molecular and biological features of TAP system. The expression levels of its protein complexes were determined by PAP immunostaining and Western blotting. Results PAP antibody immunostaining showed that MT1 MMP hTAP protein complexes (dark blue) were expressed in chick fibroblast cells. Western blot analysis of its protein complexes using MT1 MMP specific antibody showed a prominent band (68?10 3) of mobility consistent with fully processed MT1 MMP hTAP after being cleaved by the TEV proteinase. Conclusion The construction of TAP system in chick fibroblast cells and expression of MT1 MMP hTAP protein complexes will be helpful for the studies of the MT1 MMP hTAP interaction proteins in the human tumor cells.

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

The Effect of the Genistein on the Proliferation of HT1080 and Expression of Membrane Type 1-Matrix Metalloproteinase (MT1-MMP) mRNA

Matrix metalloproteinases have long been viewed as ideal candidates for proteinases that enables tumor cells to permeated basement membrane defenses and invade surrounding tissue. There is growing evidence that the MMPs have an expanded role, as they are important for the creation and maintenance of a microenvironment that facilitates growth and angiogenesis of tumors at primary and metastatic sites. MT-MMPs are not secreted but instead remaining attached to cell surfaces. Although not all of the MT-MMPs are fully characterized, MT-MMPs have important role in localizing and activating secreted MMPs. The MMP genes are transcriptionally responsive to a wide variety of oncogene, growth factors, cytokine, and hormones. Currently, a number of MMP inhibitors are being developed and some have reached clinical trials as anti-metastatic or anti-cancer therapies. MT1-MMP is involved in the activation of proMMP-2. MT1-MMP is significant not only as a tumor marker but as a new target for chemotherapy against cancer. The purpose of this study was to evaluate the effects of protein kinase C inhibitor(genistein) on the proliferation of HT1080 and expression of MT1-MMP mRNA. Human fibrosarcoma cell line HT1080 was cultured and divided 2 groups. The experimental group was treated with 100 microM genistein and incubated 12h, 24h for [3H]-thymidine uptake assay and northern hybridization individually. And the control group was treated with same amount of PBS for the above procedures. [3H]-thymidine incorporation was measured with beta ray detector. And RT-PCR and northern blotting for MT1-MMP mRNA was performed. The results were as follows 1. [3H]-thymidine uptake was reduced in experimental group with statistical significance. 2. MT1-MMP mRNA expression was significantly reduced in experimental group. These results showed that protein kinase C inhibitor (genistein) inhibited proliferation of HT1080 and almost completely blocked transcription of MT1-MMP mRNA. So, it is possible to use the protein kinase inhibitor (genistein) as anti-metastatic and anti-proliferative agent.

Matrix metalloproteinases in human gliomas: activation of matrix metalloproteinase-2 (MMP-2) may be correlated with membrane-type-1 matrix metalloproteinase (MT1-MMP) expression

To evaluate possible roles of matrix metalloproteinase (MMP)-1, -2, tissue inhibitor of metalloproteinase (TIMP)-1, -2 and membrane-type-1 matrix metalloproteinase (MT1-MMP) in invasion of human gliomas, expressions of these proteins were investigated in ten cases of human glioma and two meningioma tissues and eight human glioma cell lines. In gelatin zymography, MMP-2 activities of glioblastomas were higher than astrocytomas. The activated form of MMP-2 was seen in five of six cases of glioblastomas, but not in astrocytomas. MMP-9 activity was detected in all cases of malignant astrocytomas but the reactivity of MMP-9 was weaker than that of MMP-2. MT1-MMP mRNA expression in glioblastomas was higher than that in astrocytomas. Five cases of glioblastomas with activated form of MMP-2 had MT1-MMP expressions. In vitro, human glioma cell lines with high expression of MT1-MMP also showed high MMP-2 activity. TIMP-1 transcripts were constitutively present in almost all glioma tissues and cell lines, whereas TIMP-2 mRNA were weak especially in malignant gliomas. Imbalance of TIMP-2/MMP-2 was observed using immunoprecipitation analysis in a glioma cell line. High expression of MMP-2 and MT1-MMP is possibly involved in invasiveness of malignant glioma.

Matrix metalloproteinases in human gliomas: activation of matrix metalloproteinase-2 (MMP-2) may be correlated with membrane-type-1 matrix metalloproteinase (MT1-MMP) expression

To evaluate possible roles of matrix metalloproteinase (MMP)-1, -2, tissue inhibitor of metalloproteinase (TIMP)-1, -2 and membrane-type-1 matrix metalloproteinase (MT1-MMP) in invasion of human gliomas, expressions of these proteins were investigated in ten cases of human glioma and two meningioma tissues and eight human glioma cell lines. In gelatin zymography, MMP-2 activities of glioblastomas were higher than astrocytomas. The activated form of MMP-2 was seen in five of six cases of glioblastomas, but not in astrocytomas. MMP-9 activity was detected in all cases of malignant astrocytomas but the reactivity of MMP-9 was weaker than that of MMP-2. MT1-MMP mRNA expression in glioblastomas was higher than that in astrocytomas. Five cases of glioblastomas with activated form of MMP-2 had MT1-MMP expressions. In vitro, human glioma cell lines with high expression of MT1-MMP also showed high MMP-2 activity. TIMP-1 transcripts were constitutively present in almost all glioma tissues and cell lines, whereas TIMP-2 mRNA were weak especially in malignant gliomas. Imbalance of TIMP-2/MMP-2 was observed using immunoprecipitation analysis in a glioma cell line. High expression of MMP-2 and MT1-MMP is possibly involved in invasiveness of malignant glioma.

Membrane activator of the 72 kDa type IV collagenase in malignant breast carcinoma patients: Expression of membrane-type 1-matrix metalloproteinase (MT1-MMP)

In this study, we determined proMMP-2 activating capacity of membrane extract prepared from the tissue of invasive ductal carcinoma of breast by zymogram gel analysis. We compared the effect of membrane extract on the activation of the latent type IV collagenases with that of the organic mercurial compound leg, APMA)-induced self cleavage of the latent type IV collagenases. We also compared the expression levels of MT1-MMP between invasive carcinoma and normal tissue by Western blot, Northern blot and semi-quantitative RT-PCR analysis. Our result demonstrated that the specificity of processing by breast carcinoma membrane activator corresponds to the specificity of MT1-MMP, which clearly showed the conversion of 72-kDa proMMP-2 to the activated form while APMA processed both 72- and 92-kDa proMMPs to their activated forms. MT1-MMP protein and mRNA were expressed both in invasive carcinoma and normal tissues, and the expression levels in both tissues were comparable. Quantitative analysis of the mRNA level by RT-PCR revealed that the difference of MT1-MMP mRNA between carcinomas and normal tissues was not statistically significant on Wilcoxon signed-ranks test (P>0.05). The results from the study on the expression of MT1-MMP gene suggest that the cellular activation of MMP-2 in breast tissue, requires additional effects in addition to up-regulation of MT1-MMP.