The biochemical properties of muscle extracellular matrix are essential for stem cell adhesion, motility, proliferation and myogenic development. Recombinant elastin-like polypeptides are synthetic polypeptides that, besides maintaining some properties of the native protein, can be tailored by fusing bioactive sequences to their C-terminal. Our laboratory synthesized several Human Elastin-Like Polypeptides (HELP) derived from the sequence of human tropoelastin. Here, we developed a novel HELP family member by fusing the elastin-like backbone to the sequence of human Epidermal Growth Factor. We employed this synthetic protein, named HEGF, either alone or in combination with other proteins of the HELP family carrying RGD-integrin binding sites, as adhesion substrate for C2C12 myoblasts and satellite cells primary cultures. Adhesion of myoblasts to HEGF-based substrates induced scattering, decreased adhesion and cytoskeleton assembly; the concomitant presence of the RGD motifs potentiated all these effects. Recombinant substrates induced myoblasts proliferation, differentiation and the development of multinucleated myotubes, thus favoring myoblasts expansion and preserving their myogenic potential. The effects induced by adhesion substrates were inhibited by AG82 (Tyrphostin 25) and herbimycin A, indicating their dependence on the activation of both the EGF receptor and the tyrosine kinase c-src. Finally, HEGF increased the number of muscle stem cells (satellite cells) derived from isolated muscle fibers in culture, thus highlighting its potential as a novel substrate for skeletal muscle regeneration strategies.
- MeSH
- buněčná adheze fyziologie MeSH
- buněčná diferenciace fyziologie MeSH
- epidermální růstový faktor metabolismus fyziologie MeSH
- extracelulární matrix MeSH
- kmenové buňky cytologie MeSH
- kosterní svalová vlákna cytologie MeSH
- kosterní svaly cytologie MeSH
- kultivované buňky MeSH
- myoblasty cytologie MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- pohyb buněk fyziologie MeSH
- primární buněčná kultura MeSH
- proliferace buněk fyziologie MeSH
- satelitní buňky kosterního svalu metabolismus fyziologie MeSH
- signální transdukce MeSH
- vývoj svalů fyziologie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: The gonadotropin-induced resumption of oocyte meiosis in preovulatory follicles is preceded by expression of epidermal growth factor (EGF)-like peptides, amphiregulin (AREG) and epiregulin (EREG), in mural granulosa and cumulus cells. Both the gonadotropins and the EGF-like peptides possess the capacity to stimulate resumption of oocyte meiosis in vitro via activation of a broad signaling network in cumulus cells. To better understand the rapid genomic actions of gonadotropins (FSH) and EGF-like peptides, we analyzed transcriptomes of cumulus cells at 3 h after their stimulation. METHODS: We hybridized aRNA from cumulus cells to a pig oligonucleotide microarray and compared the transcriptomes of FSH- and AREG/EREG-stimulated cumulus cells with untreated control cells and vice versa. The identified over- and underexpressed genes were subjected to functional genomic analysis according to their molecular and cellular functions. The expression pattern of 50 selected genes with a known or potential function in ovarian development was verified by real-time qRT-PCR. RESULTS: Both FSH and AREG/EREG increased the expression of genes associated with regulation of cell proliferation, cell migration, blood coagulation and extracellular matrix remodeling. FSH alone induced the expression of genes involved in inflammatory response and in the response to reactive oxygen species. Moreover, FSH stimulated the expression of genes closely related to some ovulatory events either exclusively or significantly more than AREG/EREG (AREG, ADAMTS1, HAS2, TNFAIP6, PLAUR, PLAT, and HSD17B7). In contrast to AREG/EREG, FSH also increased the expression of genes coding for key transcription factors (CEBPB, FOS, ID1/3, and NR5A2), which may contribute to the differing expression profiles of FSH- and AREG/EREG-treated cumulus cells. CONCLUSIONS: The impact of FSH on cumulus cell gene transcription was higher than the impact of EGF-like factors in terms of the number of cell functions affected as well as the number of over- and underexpressed genes. Both FSH and EGF-like factors overexpressed genes involved in the post-ovulatory switch in steroidogenesis and tissue remodelling. However, FSH was remarkably more efficient in the up-regulation of several specific genes essential for ovulation of matured oocytes and also genes that been reported to play an important role in maturation of cumulus-enclosed oocytes in vitro.
- MeSH
- amfiregulin farmakologie fyziologie MeSH
- epidermální růstový faktor farmakologie fyziologie MeSH
- epiregulin farmakologie fyziologie MeSH
- folikuly stimulující hormon farmakologie fyziologie MeSH
- kultivované buňky MeSH
- kumulární buňky účinky léků metabolismus MeSH
- oocyty účinky léků metabolismus MeSH
- prasata MeSH
- regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Podle posledních dostupných údajů z roku 2005 v České republice bronchogenní karcinom vykazuje u mužů nejvyšší incidenci (93/100 000) ze všech maligních onemocnění a současně je u mužů nejčastější příčinou úmrtí na zhoubné nádory (mortalita u mužů je 90/100 000). U žen se v roce 2005 s incidencí 30/100 000 karcinom plic dostal na 3. místo. Celková incidence v roce 2005 byla v České republice 60/100 000. Mezi zeměmi, které shromažďují údaje o karcinomu plic, byla v roce 2002 Česká republika na 7. místě s incidencí 65/100 000, a Slovensko dokonce na 5. místě s incidencí 67/100 000. U žen pozorujeme trvale vzestupný trend v počtu nově zjištěných onemocnění. V celosvětovém měřítku zaujímá bronchogenní karcinom mezi všemi zhoubnými nádory druhé místo.
- Klíčová slova
- Herceptin, Erbitux, Iressa, Tarceva, Avastin, vandetanib, Zactima,
- MeSH
- bevacizumab MeSH
- biologická terapie trendy využití MeSH
- cetuximab MeSH
- epidermální růstový faktor fyziologie MeSH
- erbB receptory antagonisté a inhibitory MeSH
- erlotinib MeSH
- gefitinib MeSH
- hodnocení léčiv MeSH
- lidé MeSH
- monoklonální protilátky farmakologie MeSH
- nemalobuněčný karcinom plic farmakoterapie terapie MeSH
- receptor 2 pro vaskulární endoteliální růstový faktor antagonisté a inhibitory MeSH
- trastuzumab MeSH
- tyrosinkinasy antagonisté a inhibitory MeSH
- vaskulární endoteliální růstové faktory fyziologie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
BACKGROUND: Autologous dermal fibroblasts may be useful in the treatment of skin wounds and for the enhancement of keratinocyte proliferation. This paper addressed the following questions: (1) can cultured fibroblasts (CF) be transplanted as suspensions to full-thickness skin wounds and do they influence wound healing; (2) will the transplanted CF be integrated into the new dermis; (3) can a transgene that encodes a secretable marker, human epidermal growth factor (hEGF), be expressed in the wound fluid by the transplanted CF; and (4) do CF cotransplanted with cultured keratinocytes (CK) influence the rate of wound healing? METHODS: Suspensions of CF were transplanted alone or together with CK to full-thickness wounds covered with liquid-containing chambers in an established porcine model. RESULTS: Transplantation of CF accelerated reepithelialization as determined from wound histologies and sequential measurements of protein efflux over the wound surface. CF transfected with a marker gene, beta-galactosidase, resulted in in vivo gene expression and demonstrated that transplanted CF integrated into the developing dermis. Transplantation of hEGF gene-transfected CF resulted in significant hEGF expression in wound fluid. The hEGF levels peaked at day 1 (2450 pg/ml) and then sharply decreased to low levels on day 6. CF cotransplanted with CK led to greater number of keratinocyte colonies in the wound and accelerated reepithelialization as compared with CK alone. CONCLUSIONS: Transplanted CF integrated into the dermis, accelerated reepithelialization, and improved the outcome of CK transplantation. CF may also be used for the expression of transgenes in wound and wound fluid.
- MeSH
- autologní transplantace MeSH
- epidermální růstový faktor fyziologie MeSH
- fibroblasty transplantace MeSH
- hojení ran * MeSH
- keratinocyty transplantace MeSH
- kultivované buňky MeSH
- kůže cytologie MeSH
- prasata MeSH
- suspenze MeSH
- transgeny MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Research Support, U.S. Gov't, P.H.S. MeSH
- MeSH
- biologické faktory fyziologie MeSH
- blokátory kalciových kanálů farmakologie MeSH
- epidermální růstový faktor farmakologie fyziologie MeSH
- granulocyty fyziologie sekrece MeSH
- ovariální folikul fyziologie MeSH
- prasata MeSH
- progesteron biosyntéza sekrece MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
