A 170-kDa membrane-bound protease is associated with the expression of invasiveness by human malignant melanoma cells.

Aoyama, Atsuko; Chen, Wen-Tien

doi:10.1073/pnas.87.21.8296

Cited by 112 publications

(99 citation statements)

References 32 publications

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“…Our and other researchers' experiments [24] show that the phenotype of normal dermal fibroblasts cultured in vitro resembles a phenotype more like reactive fibroblasts of healing wounds than fibroblasts in healthy skin where FAP-α expression is not observed [21,[25][26][27]. We suspect that the effect of tranilast on keloid tissue degradation might result from its influence on enzymatic activity of fibroblasts surrounding keloids but not keloid fibroblasts.…”

Section: Suzawa Et Al Revealed That Tranilast Selective Inhibition Omentioning

confidence: 65%

The effect of tranilast on fibroblast activation protein α (FAP-α) expression in normal and keloid fibroblasts in vitro

Antończak¹,

Jurzak²,

Adamczyk³

et al. 2017

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Introduction. Tranilast (N-(3',4'-demethoxycinnamoyl)-anthranilic acid)is an anti-allergic drug. Its mechanism of action is based on the inhibition of antigen-induced release of chemical mediators from mast cells and basophils. It also reveals antifibroproliferative activities. These properties of tranilast are used in the treatment of hypertrophic scars and keloids. Keloids are characterized by incorrect extracellular matrix components turnover. Fibroblasts derived from keloids reveal overproduction of collagen type I and decreased degradation of extracellular matrix in comparison with normal fibroblasts. Fibroblast activation protein α (FAP-α) may play an important role in remodeling of extracellular matrix and the invasive properties of keloids. Objective. In the present study, the effect of tranilast on expression of FAP-α gene and its protein was evaluated in normal human dermal fibroblasts and fibroblasts derived from keloids cultured in vitro. Materials and methods. In the first stage of the study, the influence of tranilast on cell viability was estimated. The second stage of the study included the quantitative evaluation of FAP-α mRNA expression in normal and keloid fibroblasts treated with tranilast. The third stage of the study comprised fibroblast activation protein α expression analysis in the examined cells treated with tranilast. Results and conclusions. The expression of FAP-α gene and fibroblast activation protein α is higher in keloid fibroblasts. Tranilast at concentrations of 3 μM and 30 μM up-regulated mRNA FAP-α expression in normal fibroblasts but did not influence keloid fibroblasts. The drug, at concentrations of 30 μM and 300 μM up-regulated fibroblast activation protein α expression in normal fibroblasts and did not influence keloid fibroblasts. Tranilast antiproliferative effect is not associated with FAP-α expression in keloid fibroblasts. Original article/Praca oryginalnaThe effect of tranilast on fibroblast activation protein α (FAP-α) expression in normal and keloid fibroblasts in vitro Wpływ tranilastu na ekspresję białka aktywującego fibroblasty α (FAP-α) w fibroblastach prawidłowych oraz fibroblastach keloidowych in vitro

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Section: Suzawa Et Al Revealed That Tranilast Selective Inhibition Omentioning

confidence: 65%

The effect of tranilast on fibroblast activation protein α (FAP-α) expression in normal and keloid fibroblasts in vitro

Antończak¹,

Jurzak²,

Adamczyk³

et al. 2017

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“…The gel was prepared by incorporating the protein substrate of interest (gelatin) within the polymerised acrylamide matrix. The enzyme sample was resolved by 10% native PAGE gel in the presence of 1mg/ml gelatin (denatured type 1 collagen; Sigma) (Aoyama & Chen, 1990). The method of Laemmli was followed, excluding any reducing agents or boiling procedures.…”

Section: Gelatin Zymographymentioning

confidence: 99%

Purification, identification and characterisation of seprase from bovine serum

Collins

McMahon

O’Brien

et al. 2004

The International Journal of Biochemistry & Cell Biology

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ABBREVIATIONSAMC, 7-amino-4-methylcoumarin; BCA, bicinchoninic acid; CPC, calcium phosphate cellulose; DFP, diisopropylfluorophosphate; FPLC, fast protein liquid chromatography; HIC, hydrophobic interaction chromatography; PO, prolyl oligopeptidase; TFA, trifluoroacetic acid; Z, N-benzyloxycarbonyl; ZIP, Z-Pro-prolinal-insensitive Z-Gly-Pro-AMC-hydrolysing peptidase. ABSTRACTThe study and identification for the first time of a soluble form of a Seprase activity from bovine serum is presented. To date, this activity has only been reported to be an integral membrane protease but has been known to shed from its membrane. The activity was purified 30,197-fold to homogeneity, using a combination of column chromatographies, from bovine serum. Inhibition by DFP, resulting in an IC 50 of 100nM confirms classification as a serine protease. The protease after separation and visualisation by native PAGE was subjected to tryptic digestion and the subsequent peptides sequenced. Each peptide sequenced was found to be present in the primary structure of Seprase/Fibroblast Activation Protein (FAP), a serine gelatinase specific for proline-containing peptides and macromolecules. Substrate specificity studies using kinetic, RP-HPLC and LC-MS analysis of synthetic peptides suggest that this peptidase has an extended substrate-binding region in addition to the primary specificity site S 1 . This analysis revealed at least five subsites to be involved in enzyme-substrate binding, 2 with the smallest peptide cleaved being a tetrapeptide. A proline residue in position P 1 was absolutely necessary therefore showing high primary substrate specificity for the Pro-X bond, while a preference for a hydrophobic residue at the C-terminal end of the scissile bond (P 1 ʹ′) was evident. The enzyme also showed complete insensitivity to the prolyl oligopeptidase specific inhibitors, JTP-4819, Fmoc-Ala-pyrrCN and Z-Phe-Pro-BT. To date, no physiological substrate has clearly been defined for this protease but its ability to effectively degrade gelatin suggests a candidate protein substrate in vivo and a possible role in extracellular matrix protein degradation.

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“…These disease processes are mediated in part by the aberrantly high cell surface protease activities of breast cancer cells. Seprase, also called fibroblast activation protein-␣, is a cell surface protease that is overexpressed in breast cancer (1,2) and other cancers such as melanoma (3). Moreover, tumor tissues from breast cancer patients have aberrantly high levels of seprase activity (4).…”

Section: Introductionmentioning

confidence: 99%

“…Seprase has long been implicated in promoting tumor cell invasion (5-7). Indeed, investigation of melanoma cell lines revealed that those with high levels of seprase were most invasive (3,5,8). Moreover, sepraseexpressing melanoma cells degrade extracellular matrix more extensively than cells with low levels of seprase (5).…”

Section: Introductionmentioning

confidence: 99%

Seprase Promotes Rapid Tumor Growth and Increased Microvessel Density in a Mouse Model of Human Breast Cancer

Huang

Wang²,

Kelly³

2004

Cancer Research

109

113

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Seprase is a cell surface serine protease that is expressed to high levels by invading human breast carcinoma cells. To investigate the role of seprase in breast cancer, MDA MB-231 human mammary adenocarcinoma cells were engineered to express active seprase to high levels. All cells grow rapidly in cell culture. But differences are discovered when the cells are tested for tumorigenicity, growth, and microvessel density by implantation into the mammary fat pads of female severe combined immunodeficient mice. Control transfectants that do not express seprase grow slowly whereas cells that express seprase to high levels form fastgrowing tumors that are highly vascular. Microvessel density is elevated in tumors of two different lines of seprase transfectants to 146 ؎ 67.4 and 144 ؎ 33.42 vessels/mm 2 as compared with 50.5 ؎ 12.9 vessels/mm 2 for tumors of control-transfected cells that do not express seprase. Sepraseexpressing cells are better able to attract blood vessels and exhibit rapid tumor growth.

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A 170-kDa membrane-bound protease is associated with the expression of invasiveness by human malignant melanoma cells.

Cited by 112 publications

References 32 publications

The effect of tranilast on fibroblast activation protein α (FAP-α) expression in normal and keloid fibroblasts in vitro

The effect of tranilast on fibroblast activation protein α (FAP-α) expression in normal and keloid fibroblasts in vitro

Purification, identification and characterisation of seprase from bovine serum

Seprase Promotes Rapid Tumor Growth and Increased Microvessel Density in a Mouse Model of Human Breast Cancer

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