Staining Of MMP-7 Research Articles

Cell culture model predicts human disease: Altered expression of junction proteins and matrix metalloproteinases in cervical dysplasia

BackgroundCervical cancer is necessarily caused by human papillomaviruses, which encode three oncogenes manifesting their functions by interfering with a number of cellular proteins and pathways: the E5, E6, and E7 proteins. We have earlier found in our microarray studies that the E5 oncogene crucially affects the expression of cellular genes involved in adhesion and motility of epithelial cells.MethodsIn order to biologically validate our previous experimental findings we performed immunohistochemical staining of a representative set of tissue samples from different grades of high-risk human papillomavirus associated cervical disease as well as normal squamous and columnar cervical epithelium. Three-dimensional collagen raft cultures established from E5-expressing and control epithelial cells were also examined. The expression of p16, matrix metalloproteinase (MMP) -7, MMP-16, cytokeratin (CK) 8/18, laminin, E-cadherin and beta-catenin was studied.ResultsIn agreement with our previous microarray studies, we found intense staining for E-cadherin and beta-catenin in adherens junctions even in high-grade cervical lesions. Staining for MMP-16 was increased in severe disease as well. No significant change in staining for MMP-7 and cytokeratin 8/18 along with the grade of cervical squamous epithelial disease was observed.ConclusionsHere we have confirmed, using tissue material from human papillomavirus associated lesions, some of the cellular gene expression modifications that we earlier reported in an experimental system studying specifically the E5 oncogene of papillomaviruses. These findings were partially surprising in the context of cervical carcinogenesis and emphasize that the complexity of carcinogenesis is not yet fully understood. Microarray approaches provide a wide overwiev of gene expression in experimental settings, which may yield biologically valid biomarkers for disease diagnostics, prognosis, and follow-up.

Detection of Colorectal Adenomas Using a Bioactivatable Probe Specific for Matrix Metalloproteinase Activity

A significant proportion of colorectal adenomas, in particular those that lack an elevated growth component, continue to escape detection during endoscopic surveillance. Elevation of the activity of matrix metalloproteinases (MMPs), a large family of zinc endopeptidases, in adenomas serves as a biomarker of early tumorigenesis. The goal of this study was to assess the feasibility of using a newly developed near-infrared bioactivatable probe (MMPSense 680) that reports the activity of a broad array of MMP isoforms to detect early colorectal adenomas. Adenomatous polyposis coli (Apc)+/Min-FCCC mice that spontaneously develop multiple colorectal adenomas were injected with MMPSense 680, and the colons were imaged in an IVIS Spectrum system ex vivo. Image analyses were correlated with histopathologic findings for all regions of interest (ROIs). The biochemical basis of fluorescent signal was investigated by immunohistochemical staining of MMP-7 and -9. A strong correlation (Kendall = 0.80) was observed between a positive signal and the presence of pathologically confirmed colonic adenomas; 92.9% of the 350 ROIs evaluated were classified correctly. The correlation between two independent observers was 0.87. MMP-7 expression was localized to epithelial cells of adenomas and microadenomas, whereas staining of MMP-9 was found in infiltrating polymorphonuclear leukocytes within the adenomas. MMPSense 680 identifies colorectal adenomas, both polypoid and nonpolypoid, in Apc+/Min-FCCC mice with high specificity. Use of this fluorescent probe in combination with colonoscopy could aid in preventing colorectal neoplasias by providing new opportunities for early detection and therapeutic intervention.

Immunohistochemical expression of matrilysins (MMP-7 and MMP-26) in ameloblastomas and adenomatoid odontogenic tumors

The aim was to evaluate the expression of matrix metalloproteinases (MMPs) 7 and 26 in ameloblastomas and adenomatoid odontogenic tumors (AOTs). Twenty intraosseous solid ameloblastomas and 10 intraosseous AOTs were evaluated regarding immunohistochemical expression of MMP-7 and -26 in the epithelium and stroma. There was no statistically significant difference in MMP-7 and -26 expression between the epithelium of ameloblastomas (P = .50) and AOTs (P = .90). Stromal staining for MMP-7 was evident in all cases. For MMP-26, stromal staining was observed in 65% of ameloblastomas and 50% of AOTs, and this difference was not statistically significant (P = .69). The marked expression of these matrilysins suggests their role in the process of tissue remodeling and growth in the studied tumors, but it does not relate to the their distinct patterns of aggressiveness.

Tissue diffusion and retention of metalloproteinases in ascending aortic aneurysms and dissections

Histopathological alterations in human aneurysms and dissections of the thoracic ascending aorta include areas of mucoid degeneration within the medial layer, colocalized with areas of cell disappearance and disruption of extracellular matrix elastic and collagen fibers. We studied the presence of matrix metalloproteinases in relation to their capacity to diffuse through the tissue or to be retained in areas of mucoid degeneration in aneurysms and dissections of the ascending aorta. Ascending aortas from 9 controls, 33 patients with aneurysms, and 14 with acute dissections, all collected at surgery, were analyzed. The morphological aspect was similar whatever the etiology or phenotypic expression of the pathological aortas, involving areas of extracellular matrix breakdown and cell rarefaction associated with mucoid degeneration. Release of proMMP-2, constitutively expressed by smooth muscle cells, was not different between controls and aneurysmal aortas, whereas the aneurysmal aortas released more of the active form. Release of pro and active MMP-9 was also similar between controls and aneurysmal aortas. Immunohistochemical staining of MMP-2 and MMP-9 was weak in both control and pathological aortas. In contrast, released MMP-7 (matrilysin) and MMP-3 (stromelysin-1) could not be detected in conditioned media but were present in tissue extracts with no detectable quantitative difference between controls and pathological aortas. Immunohistochemical staining of MMP-7 and MMP-3 revealed their retention in areas of mucoid degeneration, and semiquantitative evaluation of immunostaining showed more MMP-7 in pathological aortas than in controls. In conclusion, areas of mucoid degeneration, the hallmark of aneurysms, and dissections of thoracic ascending aortas, whatever their etiology, are not inert and can retain specific proteases.

Alterations of Extracellular Matrix Components and Proteinases in Human Corneal Buttons With INTACS for Post-Laser In Situ Keratomileusis Keratectasia and Keratoconus

To perform an immunohistochemical evaluation of corneas with INTACS for post-laser in situ keratomileusis (LASIK) keratectasia and keratoconus, obtained after corneal transplantation. Corneas from 1 patient with INTACS for post-LASIK keratectasia and 2 patients with INTACS for keratoconus were obtained within 3 hours after penetrating keratoplasty, and cryostat sections were analyzed by immunostaining for 35 extracellular matrix (ECM) components and proteinases. In the stroma of all corneas next to an INTACS implant, ECM components typically associated with fibrosis were observed. These included tenascin-C, fibrillin-1, and types III, IV (alpha1/alpha2 chains), and XIV collagen. Also, significant deposition of perlecan, nidogen-2, and cellular fibronectin was revealed in the same locations. The keratoconus cases displayed typical Bowman layer breaks and subepithelial fibrosis with deposition of various ECM components. In all cases, some keratocytes around INTACS were positive for specific proteinases associated with stromal remodeling, including cathepsins F and H, matrix metalloproteinase (MMP)-1, MMP-3, and MMP-10. Staining for MMP-7 was variable; MMP-2 and MMP-9 were mostly negative. Patterns of type IV collagen alpha 3, alpha 4, and alpha 6 chains; types VI and VIII collagen; laminin-332, alpha 4, alpha 5, beta1, beta2, and gamma 1 laminin chains; vitronectin; thrombospondin-1; urokinase; EMMPRIN; and cathepsins B and L were unchanged around INTACS in all 3 cases compared with normal. Abnormal accumulation of fibrotic ECM components and proteinases near INTACS suggests ongoing lysis and remodeling of corneal stroma. Specific changes observed in each case may be related to underlying pathology.

Role of Matrix Metalloproteinase-7 in Colorectal Adenomas

Matrix metalloproteinases (MMPs) degregade and remodel the extracellular matrix. They are known to be overexpressed as normal mucosa progresses to adenomas and carcinomas. In our prospective study we measured the overexpression of MMP-7 immunohistochemically in various types of colonic adenomas. Although MMP-7 has already been shown to be overexpressed in various types of colonic adenomas, tubular versus villous adenomas had not been further seperated to date. Seventy-six patients had either normal mucosa (n=15) or tubular (n=32), tubulovillous (n=16), or villous (n=13) colonic adenoma. MMP-7 expression was classified into three categories, as negative, weakly stained, or strongly stained, depending on the percentage of cells stained. Each adenoma was graded according to the percentage of strongly stained areas in the adenoma as G0, G1, G2, or G3. Sixty-nine percent of villous adenomas showed grade 3 staining of MMP-7, versus none of the tubular adenomas. G0 and G1 staining was not detected in the villous adenomas. The results of the study show that the degrees of overexpression of the three subtypes of colonic adenomas were statistically significantly different. In conclusion, MMP-7 overexpression is thought to be an early event in the adenoma-carcinoma pathway.

Expression of matrix metalloproteinases 7 and 9 in non-small cell lung cancer: Relation to clinicopathological factors, β-catenin and prognosis

Matrix metalloproteinases (MMPs) have been shown to have a significant role in determining cancer cell behaviour. The present study was undertaken to analyze the expression and prognostic value of MMP-7 and MMP-9 in non-small cell lung cancer (NSCLC). The relationship of MMP-7 with beta-catenin was also evaluated. The study consists of 212 patients with resected NSCLC. Tumour samples were stained immunohistochemically, and the expression of MMP-7 and MMP-9 was evaluated in both tumour cells and peritumoural stromal tissue. The results were compared to clinicopathological factors of the patients. A high staining of MMP-7 and MMP-9 in tumour cells was noted in 62 (30%) and 113 (57%) cases, respectively. Expression of MMP-7 was noted more often in adenocarcinomas than in other histological types (p=0.022). High cancer cell associated MMP-7 was related to lower T-factor (p=0.037), better tumour differentiation (p=0.005) and normal beta-catenin expression in tumour cells (p=0.001). A high MMP-9 expression in tumour cells was related to poor tumour differentiation (p=0.016). The stromal signal for MMP-9 was observed in 58 (32%) cases and was linked with higher tumour grade (p=0.031). In survival analyses the significant predictors of survival were histological type of tumour and tumour stage (p=0.0009 and 0.0012, respectively). MMP-7 or MMP-9 signals were not related to patient's outcome. The results show that high MMP-9 expression indicates aggressive, and high MMP-7 less aggressive tumour behaviour in NSCLC. However, MMP-7 and MMP-9 expressions had no prognostic value in NSCLC.

Matrilsin expression is a useful marker of submucosal invasion and lymph node metastasis in early stage signet ring cell carcinoma of the stomach

Matrilysin (MMP-7) is considered to play an important role in tumor progression and metastasis. The aim of this study was to examine the MMP-7 expression of early-stage undifferentiated gastric carcinoma, and to investigate differences between gastric signet ring cell (SIG) and other undifferentiated carcinomas (non-SIG). Immunohistochemical staining of MMP-7 was performed using specimens from 150 patients with early-stage gastric undifferentiated carcinomas (76 SIG, 74 non-SIG). SIG had a larger proportion of mucosal-confined carcinoma and a lower rate of lymphatic invasion than non-SIG (P < 0.05). The incidence of the positive expression of MMP-7 in submucosal SIG was significantly higher than that of mucosal SIG (P < 0.01). In contrast, MMP-7 expression was frequently found in mucosal non-SIG, suggesting an apparent difference in the invasiveness between mucosal SIG and non-SIG. The larger the size of the mucosal SIG, the more frequently MMP-7 positive expression was demonstrated (P < 0.05). There was a significant correlation between MMP-7 positive expression and lymph node metastasis of early SIG (P < 0.05). Early SIG revealed less invasiveness than non-SIG in terms of clinicopathologic features and MMP-7 expression. Preoperative estimation of the MMP-7 expression might be useful as a predictor of submucosal invasion and lymph node metastasis in early SIG.

Mid-dermal elastolysis: A clinical, histologic, and immunohistochemical study of 11 patients

Background: Mid-dermal elastolysis is a rare entity defined by the selective loss of elastic tissue in the mid dermis. Many cases appear induced or aggravated by ultraviolet (UV) light exposure. Pathogenesis is still uncertain. Objective: Our purpose was to report on the clinical and histologic features of 11 patients with mid-dermal elastolysis. Moreover, we analyzed by immunohistochemistry leukocyte subsets and expression of metalloproteinase (MMP) with the potential to degrade elastic tissue in 7 cases. Results: All patients were women with a mean age of 31.4 years. Disease duration ranged from 4 months to 17 years. Affected areas included the trunk, neck, and upper aspect of limbs. Two patients also had Hashimoto's thyroiditis and uterine carcinoma, respectively, whereas 1 patient had undergone silicone mammoplasty. In all patients, disease onset was associated with intense UV light exposure. Moderate leukocyte infiltration in the dermis was observed mostly in recent lesions and was composed of CD3+ T cells and some CD68+ macrophages with a normal number of factor XIIIa+ dermal dendritic cells. Elastin, but not fibrillin-1 immunoreactivity disappeared from the mid dermis. MMP-9 was detected in epidermal keratinocytes and in the cytoplasm of large, angulated, multinucleated cells located in lesional dermis. These cells were negative for leukocyte, dendritic cell, macrophage, and T-cell markers and were absent in old lesions. Staining for MMP-7 and MMP-12 did not differ from control skin. Conclusion: Onset of mid-dermal elastolysis appears strongly associated with UV exposure, which may induce fibroblast-like cells to express MMP-9 that in turn could be involved in the degradation of elastic fibers. (J Am Acad Dermatol 2003;48:846-51.)