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David E Birk - One of the best experts on this subject based on the ideXlab platform.
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type v Collagen controls the initiation of Collagen fibril assembly
Journal of Biological Chemistry, 2004Co-Authors: Richard J. Wenstrup, Jane B Florer, Eric W Brunskill, Sheila M. Bell, Inna Chervoneva, David E BirkAbstract:Abstract Vertebrate Collagen fibrils are heterotypically composed of a quantitatively major and minor fibril Collagen. In non-cartilaginous tissues, type I Collagen accounts for the majority of the Collagen mass, and Collagen type V, the functions of which are poorly understood, is a minor component. Type V Collagen has been implicated in the regulation of fibril diameter, and we reported recently preliminary evidence that type V Collagen is required for Collagen fibril nucleation (Wenstrup, R. J., Florer, J. B., Cole, W. G., Willing, M. C., and Birk, D. E. (2004) J. Cell. Biochem. 92, 113–124). The purpose of this study was to define the roles of type V Collagen in the regulation of Collagen fibrillogenesis and matrix assembly. Mouse embryos completely deficient in pro-α1(V) chains were created by homologous recombination. The col5a1–/– animals die in early embryogenesis, at approximately embryonic day 10. The type V Collagen-deficient mice demonstrate a virtual lack of Collagen fibril formation. In contrast, the col5a1+/– animals are viable. The reduced type V Collagen content is associated with a 50% reduction in fibril number and dermal Collagen content. In addition, relatively normal, cylindrical fibrils are assembled with a second population of large, structurally abnormal Collagen fibrils. The structural properties of the abnormal matrix are decreased relative to the wild type control animals. These data indicate a central role for the evolutionary, ancient type V Collagen in the regulation of fibrillogenesis. The complete dependence of fibril formation on type V Collagen is indicative of the critical role of the latter in early fibril initiation. In addition, this fibril Collagen is important in the determination of fibril structure and matrix organization.
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type v Collagen heterotypic type i v Collagen interactions in the regulation of fibril assembly
Micron, 2001Co-Authors: David E BirkAbstract:Abstract Type V Collagen is a quantitatively minor fibrillar Collagen with a broad tissue distribution. The most common type V Collagen isoform is α1(V)2 α2(V) found in cornea. However, other isoforms exist, including an [α1(V)α2(V)α3(V)] form, an α1(V)3 homotrimer and hybrid type V/XI forms. The functional role and fibrillar organization of these isoforms is not understood. In the cornea, type V Collagen has a key role in the regulation of initial fibril assembly. Type I and type V Collagen co-assemble into heterotypic fibrils. The entire triple-helical domain of the type V Collagen molecules is buried within the fibril and type I Collagen molecules are present along the fibril surface. The retained NH2-terminal domains of the type V Collagen are exposed at the surface, extending outward through the gap zones. The molecular model of the NH2-terminal domain indicates that the short α helical region is a flexible hinge-like region allowing the peptide to project away from the major axis of the molecule; the short triple-helical regions serve as an extension through the hole zone, placing the tyrosine-rich domain at the surface. The assembly of early, immature fibril intermediates (segments) is regulated by the NH2-terminal domain of type V Collagen. These NH2-terminal domains alter accretion of Collagen molecules onto fibrils and therefore lateral growth. A critical density would favor the initiation of new fibrils rather than the continued growth of existing fibrils. Other type V Collagen isoforms are likely to have an important role in non-cornea tissues. This role may be mediated by supramolecular aggregates different from those in the corneal stroma or by an alteration of the interactions mediated by tissue-specific type V Collagen domains generated by different isoforms or aggregate structures. Presumably, the aggregate structure or specific domains are involved in the regionalization of fibril-associated macromolecules necessary for the tissue-specific regulation of later fibril growth and matrix assembly stages.
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Type V Collagen regulates the assembly of Collagen fibrils in cultures of bovine vascular smooth muscle cells.
Journal of cellular biochemistry, 2000Co-Authors: Kyriakos E. Kypreos, David E Birk, Vickery Trinkaus-randall, Daniel J. Hartmann, Gail E. SonensheinAbstract:Vascular smooth muscle cells (SMCs), the major cellular constituent of the medial layer of an artery, synthesize the majority of connective tissue proteins, including fibrillar Collagen types I, III, and V/XI. Proper Collagen synthesis and deposition, which are important for the integrity of the arterial wall, require the antioxidant vitamin C. Vitamin C serves as cofactor for the enzymes prolyl and lysyl hydroxylase, which are responsible for the proper hydroxylation of Collagen. Here, the role of type V Collagen in the assembly of Collagen fibrils in the extracellular matrix (ECM) of cultured vascular SMCs was investigated. Treatment of SMCs with vitamin C resulted in a dramatic induction in the levels of the cell-layer associated pepsin-resistant type V Collagen, whereas only a minor induction in the levels of types I and III Collagen was detected. Of note, the deposition of type V Collagen was accompanied by the formation of striated Collagen fibrils in the ECM. Immunohistochemistry demonstrated that type V Collagen, but not type I Collagen, became masked as Collagen fibrils matured. Furthermore, the relative ratio of type V to type I Collagen decreased as the ECM matured as a function of days in culture, and this decrease was accompanied by an increase in the diameter of Collagen fibrils. Together these results suggest that the masking of type V Collagen is caused by its internalization on continuous deposition of type I Collagen on the exterior of the fibril. Furthermore, they suggest that type V Collagen acts as framework for the initial assembly of Collagen molecules into heterotypic fibrils, regulating the diameter and architecture of these fibrils. J. Cell. Biochem. 80:146–155, 2000. © 2000 Wiley-Liss, Inc.
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Reduction of type V Collagen using a dominant-negative strategy alters the regulation of fibrillogenesis and results in the loss of corneal-specific fibril morphology.
The Journal of cell biology, 1996Co-Authors: Jeffrey K. Marchant, Thomas F. Linsenmayer, Rita A. Hahn, David E BirkAbstract:A number of factors have been implicated in the regulation of tissue-specific Collagen fibril diameter. Previous data suggest that assembly of heterotypic fibrils composed of two different fibrillar Collagens represents a general mechanism regulating fibril diameter. Specifically, we hypothesize that type V Collagen is required for the assembly of the small diameter fibrils observed in the cornea. To test this, we used a dominant-negative retroviral strategy to decrease the levels of type V Collagen secreted by chicken corneal fibroblasts. The chicken alpha 1(V) Collagen gene was cloned, and retroviral vectors that expressed a polycistronic mRNA encoding a truncated alpha 1(V) minigene and the reporter gene LacZ were constructed. The efficiency of viral infection was 30-40%, as determined by assaying beta-galactosidase activity. To assess the expression from the recombinant provirus, Northern analysis was performed and indicated that infected fibroblasts expressed high steady-state levels of retroviral mRNA. Infected cells synthesized the truncated alpha 1(V) protein, and this was detectable only intracellularly, in a distribution that colocalized with lysosomes. To assess endogenous alpha 1(V) protein levels, infected cell cultures were assayed, and these consistently demonstrated reductions relative to control virus-infected or uninfected cultures. Analyses of corneal fibril morphology demonstrated that the reduction in type V Collagen resulted in the assembly of large-diameter fibrils with a broad size distribution, characteristics similar to fibrils produced in connective tissues with low type V concentrations. Immunoelectron microscopy demonstrated the amino-terminal domain of type V Collagen was associated with the small-diameter fibrils, but not the large fibrils. These data indicate that type V Collagen levels regulate corneal fibril diameter and that the reduction of type V Collagen is sufficient to alter fibril assembly so that abnormally large-diameter fibrils are deposited into the matrix.
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Positive Regulation of Corneal Type V Collagen mRNA: Analysis by Chicken–Human Heterokaryon Formation
Experimental cell research, 1996Co-Authors: Thomas F. Linsenmayer, Frank Igoe, Eileen Gibney, Marion K. Gordon, David E BirkAbstract:Abstract Our previous studies have suggested that type V Collagen is at least one factor responsible for the characteristically small, uniform diameter of striated Collagen fibrils of the corneal stroma. These fibrils, which are heterotypic combinations of Collagen types I and V, contain four- to fivefold more type V Collagen than those of tendon and sclera. The latter are much larger and more heterodisperse. This high content of type V Collagen in cornea is reflected by an equally elevated content of α1(V) chain mRNA in corneal fibroblasts. Thus, the increased production of the molecule in cornea appears to be regulated at the level of transcription and/or mRNA stability. One possible explanation for this is that corneal fibroblasts contain positive regulatory factors that specifically upregulate transcription of the type V Collagen genes and/or increase their mRNA stability. To test this possibility, we have produced transient heterokaryons by fusing chicken corneal fibroblasts with two human noncorneal cell lines selected as containing little if any α1(V) mRNA. If the chicken corneal cells contain positive regulators that can act across species, these regulators should result in increased levels of the human α1(V) transcript. The results were evaluated by reverse transcript–polymerase chain reaction employing a primer pair selected for its ability specifically to amplify part of the human α1(V) mRNA. In fusions between chicken corneal fibroblasts and the human cell lines, after a lag of 10–14 h the heterokaryon-containing cultures showedde novoappearance or upregulation of human α1(V) chain mRNA, compared with that of the parental cell lines. Cultures of the mixed cell types that had not been fused showed no such upregulation, so the effect was not mediated by diffusible substances acting between the cells. Chicken tendon fibroblasts, a low producer of type V Collagen, when tested in the same assay, evoked no detectible increase in the human transcript. Thus, corneal cells do contain positive regulators for α1(V) chain mRNA, and this effect is at least somewhat cell specific.
Robert Garrone - One of the best experts on this subject based on the ideXlab platform.
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THE EVOLUTION OF FIBRILLAR CollagenS : A SEA-PEN Collagen SHARES COMMON FEATURES WITH VERTEBRATE TYPE V Collagen
Comparative Biochemistry and Physiology B, 1996Co-Authors: Emmanuelle Tillet, Jean-marie Franc, Suzanne Franc, Robert GarroneAbstract:The extracellular matrix of marine primitive invertebrates (sponges, polyps and jellyfishes) contains Collagen fibrils with narrow diameters. From various data, it has been hypothesized that these primitive Collagens could represent ancestral forms of the vertebrate minor Collagens, i.e. types V or XI. Recently, we have isolated a primitive Collagen from the soft tissues of the sea-pen Veretillum cynomorium(31). This report examines whether the sea-pen Collagen shares some features with vertebrate type V Collagen. Rotary shadowed images of acid-soluble Collagen molecules extracted from β-APN treated animals, positive staining of segmentlong-spacing crystallites precipitated from pepsinized Collagen, Western blots of the pepsinized α1 and α2 chains with antibodies to vertebrate types I, III and V Collagens, and in situ gold immunolabeling of ECM Collagen fibrils were examined. Our results showed that the tissue form of the sea-pen Collagen is a 340-nm threadlike molecule, which is close to the vertebrate type V Collagen with its voluminous terminal globular domain, the distribution of most of its polar amino-acid residues, and its antigenic properties.
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The evolution of fibrillar Collagens: a sea-pen Collagen shares common features with vertebrate type V Collagen.
Comparative Biochemistry and Physiology - Part B: Biochemistry and Molecular Biology, 1996Co-Authors: Emmanuelle Tillet, Suzanne Franc, Jm Franc, Robert GarroneAbstract:The extracellular matrix of marine primitive invertebrates (sponges, polyps and jellyfishes) contains Collagen fibrils with narrow diameters. From various data, it has been hypothesized that these primitive Collagens could represent ancestral forms of the vertebrate minor Collagens, i.e., types V or XI. Recently we have isolated a primitive Collagen from the soft tissues of the sea-pen Veretillum cynomorium. This report examines whether the sea-pen Collagen shares some features with vertebrate type V Collagen. Rotary shadowed images of acid-soluble Collagen molecules extracted from beta-APN treated animals, positive staining of segment-long-spacing crystallites precipitated from pepsinized Collagen, Western blots of the pepsinized alpha1 and alpha2 chains with antibodies to vertebrate types I, III and V Collagens, and in situ gold immunolabeling of ECM Collagen fibrils were examined. Our results showed that the tissue form of the sea-pen Collagen is a 340-nm threadlike molecule, which is close to the vertebrate type V Collagen with its voluminous terminal globular domain, the distribution of most of its polar amino-acid residues, and its antigenic properties.The extracellular matrix of marine primitive invertebrates (sponges, polyps and jellyfishes) contains Collagen fibrils with narrow diameters. From various data, it has been hypothesized that these primitive Collagens could represent ancestral forms of the vertebrate minor Collagens, i.e., types V or XI. Recently we have isolated a primitive Collagen from the soft tissues of the sea-pen Veretillum cynomorium. This report examines whether the sea-pen Collagen shares some features with vertebrate type V Collagen. Rotary shadowed images of acid-soluble Collagen molecules extracted from beta-APN treated animals, positive staining of segment-long-spacing crystallites precipitated from pepsinized Collagen, Western blots of the pepsinized alpha1 and alpha2 chains with antibodies to vertebrate types I, III and V Collagens, and in situ gold immunolabeling of ECM Collagen fibrils were examined. Our results showed that the tissue form of the sea-pen Collagen is a 340-nm threadlike molecule, which is close to the vertebrate type V Collagen with its voluminous terminal globular domain, the distribution of most of its polar amino-acid residues, and its antigenic properties.
Kozo Ohtsuki - One of the best experts on this subject based on the ideXlab platform.
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EFFECT OF SLAUGHTER METHOD ON DEGRADATION OF INTRAMUSCULAR TYPE V Collagen DURING SHORT‐TERM CHILLED STORAGE OF CHUB MACKEREL SCOMBER JAPONICUS
Journal of Food Biochemistry, 2002Co-Authors: Kenji Sato, Yasutaka Shigemura, Masashi Ando, Shugo Uratsujt, Mayumi Sato, Satoshi Mochizuki, Yasushi Nakamura, Kozo OhtsukiAbstract:The present paper demonstrates that a nonstntggling slaughter method can delay degradation of type V Collagen in meat of chub mackerel Scomber japonicus and softening of the meat during postharvest chilled storage. The fish were slaughtered by piercing a knife into nape (nonstruggling method) or by leaving on ground (struggling method) and then stored in an ice box. Sensory study revealed that the postharvest softening of the meat was moderated at 4 and 8 h by the non-struggling slaughter method in comparison with the struggling method. On the basis of the specific solubilization of type V Collagen and reduced tyrosine content in it, a cleavage of the nonhelical regions (telopeptides) of the type V Collagen occurred during the chilled storage in the fish slaughtered by the struggling method. The degradation of type V Collagen was also slower in the meat of the fish slaughtered by the nonstruggling method, which can be directly linked to the moderation of the postharvest softening.
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ISOLATION OF INTACT TYPE V Collagen FROM FISH INTRAMUSCULAR CONNECTIVE TISSUE
Journal of Food Biochemistry, 1998Co-Authors: Kenji Sato, Chiho Ohashi, Mitsumi Muraki, Hukiko Itsuda, Yoshihiro Yokoyama, Masao Kanamori, Kozo Ohtsuki, Makoto KawabataAbstract:A purification method for intact type V Collagen from fish intramuscular connective tissue was developed. Collagens were extracted with 0.5 M acetic acid from fish intramuscular connective tissue. Intact type V Collagen could not be isolated by conventional salt fractionation which has been used for isolation of pepsinized Collagens. Isolation was achieved by ion-exchange chromatography in the presence of 5 M urea. This is the first report on isolation of intact type V Collagen from muscle tissue.
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Involvement of Type V Collagen in Softening of Fish Muscle during Short-Term Chilled Storage
Journal of Agricultural and Food Chemistry, 1997Co-Authors: Kenji Sato, Masashi Ando, Yasuo Makinodan, Satoshi Kubota, Kyoko Origasa, Hiroshi Kawase, Haruhiko Toyohara, Morihiko Sakaguchi, Takayuki Nakagawa, Kozo OhtsukiAbstract:Type V Collagen became solubilized in softened sardine muscle after 1 day of chilled storage with the concomitant weakening of pericellular connective tissue induced by disintegration of thin Collagen fibrils. However, no significant changes were observed in the structure of interstitial connective tissue or biochemical properties of type I Collagen. Z disk in myofibrils showed structural changes, but no significant loss of longitudinal continuity of myofibrils was observed even at the deteriorated Z disk from the muscle destroyed by compression test. On the other hand, tiger puffer muscle did not show significant softening during the storage, with no significant changes in structure of connective tissues and biochemical properties of Collagens. These facts suggest that degradation of type V Collagen causes disintegration of the thin Collagen fibrils in pericellular connective tissue, weakening pericellular connective tissue and resulting in postharvest softening. Keywords: Collagen; postharvest storage; ...
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Type V Collagen in trout (Salmo gairdneri) muscle and its solubility change during chilled storage of muscle
Journal of Agricultural and Food Chemistry, 1991Co-Authors: Kenji Sato, Chiho Ohashi, Kozo Ohtsuki, Makoto KawabataAbstract:Post-mortem changes of types I and V Collagens in rainbow trout muscle were examined in relation to the softening of fish muscle during chilled storage. Degradation of helical regions of Collagens was not detected. On the other hand, the solubility of type V Collagen increased significantly, while that of type I Collagen did not change. These facts suggest that degradation of nonhelical regions or intermolecular cross-links occur preferentially in type V Collagen.
Kenji Sato - One of the best experts on this subject based on the ideXlab platform.
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EFFECT OF SLAUGHTER METHOD ON DEGRADATION OF INTRAMUSCULAR TYPE V Collagen DURING SHORT‐TERM CHILLED STORAGE OF CHUB MACKEREL SCOMBER JAPONICUS
Journal of Food Biochemistry, 2002Co-Authors: Kenji Sato, Yasutaka Shigemura, Masashi Ando, Shugo Uratsujt, Mayumi Sato, Satoshi Mochizuki, Yasushi Nakamura, Kozo OhtsukiAbstract:The present paper demonstrates that a nonstntggling slaughter method can delay degradation of type V Collagen in meat of chub mackerel Scomber japonicus and softening of the meat during postharvest chilled storage. The fish were slaughtered by piercing a knife into nape (nonstruggling method) or by leaving on ground (struggling method) and then stored in an ice box. Sensory study revealed that the postharvest softening of the meat was moderated at 4 and 8 h by the non-struggling slaughter method in comparison with the struggling method. On the basis of the specific solubilization of type V Collagen and reduced tyrosine content in it, a cleavage of the nonhelical regions (telopeptides) of the type V Collagen occurred during the chilled storage in the fish slaughtered by the struggling method. The degradation of type V Collagen was also slower in the meat of the fish slaughtered by the nonstruggling method, which can be directly linked to the moderation of the postharvest softening.
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ISOLATION OF INTACT TYPE V Collagen FROM FISH INTRAMUSCULAR CONNECTIVE TISSUE
Journal of Food Biochemistry, 1998Co-Authors: Kenji Sato, Chiho Ohashi, Mitsumi Muraki, Hukiko Itsuda, Yoshihiro Yokoyama, Masao Kanamori, Kozo Ohtsuki, Makoto KawabataAbstract:A purification method for intact type V Collagen from fish intramuscular connective tissue was developed. Collagens were extracted with 0.5 M acetic acid from fish intramuscular connective tissue. Intact type V Collagen could not be isolated by conventional salt fractionation which has been used for isolation of pepsinized Collagens. Isolation was achieved by ion-exchange chromatography in the presence of 5 M urea. This is the first report on isolation of intact type V Collagen from muscle tissue.
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Involvement of Type V Collagen in Softening of Fish Muscle during Short-Term Chilled Storage
Journal of Agricultural and Food Chemistry, 1997Co-Authors: Kenji Sato, Masashi Ando, Yasuo Makinodan, Satoshi Kubota, Kyoko Origasa, Hiroshi Kawase, Haruhiko Toyohara, Morihiko Sakaguchi, Takayuki Nakagawa, Kozo OhtsukiAbstract:Type V Collagen became solubilized in softened sardine muscle after 1 day of chilled storage with the concomitant weakening of pericellular connective tissue induced by disintegration of thin Collagen fibrils. However, no significant changes were observed in the structure of interstitial connective tissue or biochemical properties of type I Collagen. Z disk in myofibrils showed structural changes, but no significant loss of longitudinal continuity of myofibrils was observed even at the deteriorated Z disk from the muscle destroyed by compression test. On the other hand, tiger puffer muscle did not show significant softening during the storage, with no significant changes in structure of connective tissues and biochemical properties of Collagens. These facts suggest that degradation of type V Collagen causes disintegration of the thin Collagen fibrils in pericellular connective tissue, weakening pericellular connective tissue and resulting in postharvest softening. Keywords: Collagen; postharvest storage; ...
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Type V Collagen in trout (Salmo gairdneri) muscle and its solubility change during chilled storage of muscle
Journal of Agricultural and Food Chemistry, 1991Co-Authors: Kenji Sato, Chiho Ohashi, Kozo Ohtsuki, Makoto KawabataAbstract:Post-mortem changes of types I and V Collagens in rainbow trout muscle were examined in relation to the softening of fish muscle during chilled storage. Degradation of helical regions of Collagens was not detected. On the other hand, the solubility of type V Collagen increased significantly, while that of type I Collagen did not change. These facts suggest that degradation of nonhelical regions or intermolecular cross-links occur preferentially in type V Collagen.
Rosalia Sirchia - One of the best experts on this subject based on the ideXlab platform.
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Type V Collagen and protein kinase C η down‐regulation in 8701‐BC breast cancer cells
Molecular carcinogenesis, 2011Co-Authors: Claudio Luparello, Rosalia Sirchia, Alessandra LongoAbstract:We previously reported that ductal infiltrating carcinomas (d.i.c.) of the human breast display profound modifications of the stromal architecture, associated with anomalous Collagen composition. Among the major alterations observed in the interstitial Collagen, the relative increase of type V Collagen content was detected. When type V Collagen was used as an "in vitro" substrate for 8701-BC d.i.c. cells, it appeared able to restrain cell growth, inhibit cell motility and invasion "in vitro", and modify the expression levels of genes coding for apoptosis factors, caspases and stress response proteins. In the present paper we demonstrate that type V Collagen induces the down-regulation of protein kinase C η, an event that may be, at least in part, responsible of the previously-reported modifications of cell morphology and growth rate, and that appears to be involved in the already-observed changes of expression levels of genes encoding for anti- (Bcl-2) and pro-apoptotic factors (Bad, Dapk, Bcl-Xs) and enzymes (caspase 5 and 8).
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Type V Collagen-induced upregulation of capn2 (large subunit of m-calpain) gene expression and DNA fragmentation in 8701-BC breast cancer cells.
Biological chemistry, 2011Co-Authors: Claudio Luparello, Rosalia SirchiaAbstract:Type V Collagen is known to be over-deposited in the stroma of ductal infiltrating carcinomas of the breast. When used as a substrate, type V Collagen restrains growth and invasion, and affects gene expression of 8701-BC ductal infiltrating carcinomas cells. Here we supplement existing data by demonstrating type V Collagen dependent upregulation of capn2 gene expression in 8701-BC cells through differential display-PCR and Western blot assays. Furthermore, we suggest that our data obtained by centrifugal sedimentation and electrophoresis strongly suggest a correlation between calpain overproduction and DNA fragmentation, since the incubation with calpain inhibitor partly reverts the latter.
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Type V Collagen regulates the expression of apoptotic and stress response genes by breast cancer cells
Journal of cellular physiology, 2004Co-Authors: Claudio Luparello, Rosalia SirchiaAbstract:Type V Collagen is a "minor" component of normal human breast stroma, which is subjected to over-deposition in cases of ductal infiltrating carcinoma (DIC). We reported that, if used as a culture substrate for the DIC cell line 8701-BC, it exhibited poorly-adhesive properties and restrained the proliferative and motile behavior of the cell subpopulation able to attach onto it. Moreover, this Collagen species was able to trigger DNA fragmentation and impair survival of 8701-BC cells. In this study, we have extended our investigation with the aim to obtain further evidence that the death induced by type V Collagen was of the apoptotic type by (i) microscopic detection and quantitation of Apoptag-labeled cells, (ii) analysis of the expression levels of selected genes coding for apoptosis-linked factors, caspases, and stress-response proteins by conventional and semi-quantitative multiplex PCR, and (iii) evaluation of the extent of caspase activation by chromogenic assay. We report here that type V Collagen is able to determine an increase in the percentage of Apoptag-positive cells, to up-regulate Bcl-xS, Bad, Dap kinase, hsf-1, mthsp75, caspase-1, -5, -8, -9, and -14, whilst down-regulating Bcl-2, Bcl-xbeta, and hsp60. Treatment of cell lysates with chromogenic tetrapeptide substrates specific for caspase-1, -5, -8, and -9 demonstrated a marked increase of enzymatic activity in the presence of type V Collagen. Our data validate 8701-BC cell line as a suitable "in vitro" model for further and more detailed studies on the molecular mechanisms of the death response induced by type V Collagen on primary DIC cells.
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T47-D cells and type V Collagen: a model for the study of apoptotic gene expression by breast cancer cells
Biological chemistry, 2003Co-Authors: Claudio Luparello, Fabio David, Giovanna Campisi, Rosalia SirchiaAbstract:We have previously reported that type V Collagen is a poorly adhesive, anti-proliferative and motility-inhibitory substrate for the 8701-BC breast cancer cell line, which also triggers DNA fragmentation and impairs survival of the same cell line. In the present work we have extended to other breast cancer cell lines (T47-D, MDA-MB231, Hs578T) our investigation of type V Collagen influence on the DNA status and cell survival, also examining whether adhesion and growth of cells on this Collagen substrate could exert some effect on the expression level of selected apoptosis-related genes. We report here that, among the cell lines tested, only T47-D is responsive to the death-promoting influence of type V Collagen. In addition, the latter induces changes in gene expression by up-regulating p53, Waf-1, Cas, Dap kinase and caspases 1, -5 and -14 and down-regulating Bcl-2. Our data validate the T47-D line as a suitable in vitro model for further and more detailed studies on the molecular mechanisms of the death response induced by type V Collagen on mammary tumor cells.
