Decreased expression of ADAMTS-1 in human breast tumors stimulates migration and invasion

Abstract Background ADAMTS-1 (a disintegrin and metalloprotease with thrombospondin motifs) is a member of the ADAMTS family of metalloproteases. Here, we investigated mRNA and protein levels of ADAMTS-1 in normal and neoplastic tissues using qPCR, immunohistochemistry and immunoblot analyses, and we addressed the role of ADAMTS-1 in regulating migration, invasion and invadopodia formation in breast tumor cell lines. Results In a series of primary breast tumors, we observed variable levels of ADAMTS-1 mRNA expression but lower levels of ADAMTS-1 protein expression in human breast cancers as compared to normal tissue, with a striking decrease observed in high-malignancy cases (triple-negative for estrogen, progesterone and Her-2). This result prompted us to analyze the effect of ADAMTS-1 knockdown in breast cancer cells in vitro. MDA-MB-231 cells with depleted ADAMTS-1 expression demonstrated increased migration, invasion and invadopodia formation. The regulatory mechanisms underlying the effects of ADAMTS-1 may be related to VEGF, a growth factor involved in migration and invasion. MDA-MB-231 cells with depleted ADAMTS-1 showed increased VEGF concentrations in conditioned medium capable of inducing human endothelial cells (HUVEC) tubulogenesis. Furthermore, expression of the VEGF receptor (VEGFR2) was increased in MDA-MB-231 cells as compared to MCF7 cells. To further determine the relationship between ADAMTS-1 and VEGF regulating breast cancer cells, MDA-MB-231 cells with reduced expression of ADAMTS-1 were pretreated with a function-blocking antibody against VEGF and then tested in migration and invasion assays; both were partially rescued to control levels. Conclusions ADAMTS-1 expression was decreased in human breast tumors, and ADAMTS-1 knockdown stimulated migration, invasion and invadopodia formation in breast cancer cells in vitro. Therefore, this series of experiments suggests that VEGF is involved in the effects mediated by ADAMTS-1 in breast cancer cells.

Invasion of Ureaplasma diversum in bovine spermatozoids

Background Ureaplasma diversum has been associated with infertility in cows. In bulls, this mollicute colonizes the prepuce and distal portion of the urethra and may infect sperm cells. The aim of this study is to analyze in vitro interaction of U. diversum isolates and ATCC strains with bovine spermatozoids. The interactions were observed by confocal microscopy and the gentamycin internalization assay. Findings U. diversum were able to adhere to and invade spermatozoids after 30 min of infection. The gentamicin resistance assay confirmed the intracellularity and survival of U. diversum in bovine spermatozoids. Conclusions The intracellular nature of bovine ureaplasma identifies a new difficulty to control the reproductive of these animals.

Invasion of the dinoflagellate Ceratium furcoides (Levander) Langhans 1925 at tropical reservoir and its relation to environmental variables

Dinoflagellates of the genus Ceratium are chiefly marine but there are rare occurrences in freshwater. In this study we analyze the invasion and progressive establishment of Ceratium furcoides, an exotic species, in the Furnas Reservoir. Samples were taken at 36 points in the reservoir, during the months of March, June, September and December, 2007. Measurements of some physical and chemical variables were simultaneously performed at each site. The occurrence of C. furcoides was registered at 20 sites, with densities varying between 0.57 and 28,564,913.0 ind.m-3. Blooms of this species were recorded in points which were classified as mesotrophic, coinciding with the places receiving high amounts of untreated domestic sewage. C. furcoides density was correlated with temperature, nutrients (nitrate and nitrite) and water electric conductivity. The highest density was recorded in June when temperature was low. The presence of Ceratium furcoides in the reservoir apparently has not yet affected the reservoir water quality or other plankton communities. However, if it becomes fully established it could perhaps become a problem in the reservoir or even to spread out to other reservoirs in Rio Grande basin.

Die Rolle von Ephrin-B2 in der Invasion maligner Gliome

Das Glioblastoma multiforme zählt zu den häufigsten glialen Neoplasien des Menschen und weist zudem unter den Gliomen die höchste Malignität auf. Glioblastompatienten haben trotz aggressiver therapeutischer Ansätze eine mittlere Überlebenszeit von weniger als einem Jahr. Die diffuse Invasion in das umliegende Hirngewebe ist einer der Hauptgründe für die Rezidivbildung und die infauste Prognose von Glioblastompatienten. Neuere Untersuchungen lassen vermuten, dass die starke Invasion auch einer der Gründe für die beobachtete anti-angiogene Resistenz bei der Behandlung von Glioblastomen ist. Das bidirektionale EphB/Ephrin-B-System wurde bei der axonalen Wegfindung als Vermittler repulsiver Signale identifiziert und auch im Zusammenhang der Migration und Invasion von Zellen überprüft. In der vorliegenden Arbeit sollte daher die Funktion der bidirektionalen Eph- und Ephrin-Signaltransduktion in Bezug auf die Glioblastominvasion und Progression untersucht werden. rn Genetische und epigenetische Untersuchungen der EphB/Ephrin-B-Familie in einer Kohorte von Gliompatienten unterschiedlicher Malignitätsgrade identifizierten Ephrin-B2 als mögliches Tumorsuppressorgen. In Übereinstimmung damit führte die Inaktivierung von Ephrin-B2 in einem murinen Gliommodell zu einer verstärkten Invasion und einem erhöhtem Tumorwachstum in vivo. Dies konnte in verschiedenen Invasion-Assays in vitro bestätigt werden. Weiterhin zeigten unsere Untersuchungen, dass Ephrin-B2 transkriptionell durch das hypoxische Mikromilieu HIF-1α-vermittelt reprimiert wird. Da HIF-1α als transkriptioneller Aktivator Ephrin-B2 nicht direkt reprimieren kann, wurden potentielle HIF-1α-regulierte Repressoren untersucht, die für die Ephrin-B2 Herunterregulation verantwortlich sein könnten. Dabei wurde anhand von Ephrin-B2-Promotoranalysen und ChIP-Assays ZEB2 als HIF-1α-induzierbarer Repressor von Ephrin-B2 identifiziert. Zur Bestätigung der Hypothese, dass ZEB2 ein wichtiger Regulator der Tumorinvasion ist, wurden humane ZEB2-Knockdown-Glioblastomzellen generiert und in vitro sowie in vivo untersucht. Im Hinblick auf mögliche therapeutische Anwendungen wurden die ZEB2-Knockdown-Glioblastomzellen zusätzlich im Zusammenhang anti-Angiogenese-induzierter Invasion analysiert. Der Verlust von ZEB2 führte dabei zu einer verringerten Glioblastominvasion und Progression in einem Maus-Xenograft Modell. Die Behandlung der Tumoren mit dem anti-VEGF-Antikörper Avastin resultierte in einer stark erhöhten Invasion, die durch die Inaktivierung von ZEB2 und der dadurch reaktivierten repulsiven Signale von Ephrin-B2 wieder aufgehoben werden konnte. Zusammenfassend konnte in der vorliegenden Arbeit erstmals gezeigt werden, dass Ephrin-B2 als Tumorsuppressor in Gliomen agiert und durch verschiedene Mechanismen wie der genetischen und epigenetischen Kontrolle, aber auch der HIF-1α-vermittelten, ZEB2-abhängigen Repression inaktiviert wird. Dies resultiert in einer Blockade repulsiver Signale, so dass Tumorzellen diffus in das Parenchym und zu den Blutgefäßen migrieren können. Der in dieser Arbeit neu identifizierte Signalweg stellt ein attraktives therapeutisches Ziel zur Inhibition der Tumorzellinvasion dar und ermöglicht darüber hinaus der Ausbildung von Resistenzen gegenüber anti-angiogener Behandlung entgegenzuwirken. rn

Evolution of the sexual reproduction in Veronica (Plantaginaceae) : phylogeny, phylogeography and invasion

Die geschlechtliche Fortpflanzung ist ein universelles Merkmal und erlaubt es genetische Variation innerhalb von Blütenpflanzen zu schaffen. Die Evolution der sexuellen und reproduktiven Systeme wurde hier auf mehreren zeitlichen Ebenen, in verschiedenen Arten von Lebensraum studiert und mit fast allen möglichen Methoden im Labor, im Gewächshaus sowie im Feld untersucht. Drei Hauptteile sind in dieser Arbeit enthalten und entsprechen jeweils einem unterschiedlichen Niveau der Zeit: Gattung, Untergattung und Arten. Der erste Teil zeigt, dass die PO-Verhältnisse Untersuchungen systematisch in jeder Pflanzen-Gattung oder Untergattung untersucht werden müssen. Dieses güngstige, schnelle und leistungsstarke Werkzeug kann eine Vielzahl von Informationen über die Modi der Pflanzenreproduktion produzieren, ohne die Verwendung von teuren und langen Experimenten. Darüber hinaus könnte diese Maßnahme auch ergänzende Daten über die Taxonomie dieser untersuchten Gruppen geben. Das zweite Kapitel befasst sich mehr mit der Taxonomie der Ehrenpreis(Veronica)-Arten als die beiden anderen und zeigt, dass verschiedene Ereignisse der interspezifischen Reproduktion in einem der Hotspots der Artenvielfalt in Europa (der Balkan-Halbinsel) auftreten. Die Ergebnisse zeigen, dass morphologische und genetische Daten inkongruent sind und die Analyse der Taxonomie dieser Arten oder Unterart schwierig ist. Das letzteKapitel erzählt die Geschichte einer erfolgreichen Invasion, die während des letzten Jahrhunderts in Europa ablief trotz der Tatsache, dass die Arten obligate Fremdbefruchter sind und dass keine Samen-Produktion in der Region beobachtet wurde. Dieses Manuskript erläutert den Weg der Pflanze, um die “Baker-Regel“ zu umgehen. Diese Regel besagt, dass selbst-inkompatible Arten erfolgloser bei der Invasion neuer Lebensräume sind. Dennoch schafft es die hier untersuchte Art einen großen Teil der europäischen Rasen zu bevölkern und zeigt dabei genetische und morphologische Veränderungen auf diesem Weg.rnSchließlich wird in diesen drei verschiedenen Papieren versucht, die Verbindung zwischen der Mikro-und Makroevolution in der geschlechtlichen Fortpflanzun in Ehrenpreis (Veronica) unter Betracht verschiedener sexueller Systeme und der Stammesgeschichte, sowie der Migration zu klären.

MMP19 is upregulated during melanoma progression and increases invasion of melanoma cells

During the progression of cutaneous melanomas, matrix metalloproteinases (MMPs) facilitate the tumour cells to traverse the basement membrane and invade the dermis. In this study, we analysed the expression of MMP19 in the course of melanoma progression. Although MMP19 was absent in melanocytes and melanoma cells of early stages of melanoma development, its expression was strongly upregulated in the neighbouring keratinocytes that may facilitate the vertical outgrowth of melanoma cells. In contrast to early stages, MMP19 was upregulated during the vertical growth phase of melanoma and in metastases. The upregulation of MMP19 in melanoma of Clark levels IV and V correlates with that of MMP2 and also simultaneously with ceased expression of E-cadherin. To reveal whether MMP19 facilitates the invasion of melanomas, we examined adhesion and migratory capacity of selected melanoma cell lines. Melanoma cell lines with low expression of MMP19 exhibited increased adhesion to various substrates and lower migration in comparison with the cell line with higher expression of MMP19. Moreover, ectopic expression of MMP19 could restore the migratory capacity of melanoma cells with low endogenous level of MMP19. These results suggest that the increase of MMP19 expression hallmarks the progression of cutaneous melanoma and might augment melanoma growth by promoting the invasion of tumour cells.

Hybridization between distant lineages increases adaptive variation during a biological invasion: stickleback in Switzerland

The three-spined stickleback is a widespread Holarctic species complex that radiated from the sea into freshwaters after the retreat of the Pleistocene ice sheets. In Switzerland, sticklebacks were absent with the exception of the far northwest, but different introduced populations have expanded to occupy a wide range of habitats since the late 19th century. A well-studied adaptive phenotypic trait in sticklebacks is the number of lateral plates. With few exceptions, freshwater and marine populations in Europe are fixed for either the low plated phenotype or the fully plated phenotype, respectively. Switzerland, in contrast, harbours in close proximity the full range of phenotypic variation known from across the continent. We addressed the phylogeographic origins of Swiss sticklebacks using mitochondrial partial cytochrome b and control region sequences. We found only five different haplotypes but these originated from three distinct European regions, fixed for different plate phenotypes. These lineages occur largely in isolation at opposite ends of Switzerland, but co-occur in a large central part. Across the country, we found a strong correlation between a microsatellite linked to the high plate ectodysplasin allele and the mitochondrial haplotype from a region where the fully plated phenotype is fixed. Phylogenomic and population genomic analysis of 481 polymorphic amplified fragment length polymorphism loci indicate genetic admixture in the central part of the country. The same part of the country also carries elevated within-population phenotypic variation. We conclude that during the recent invasive range expansion of sticklebacks in Switzerland, adaptive and neutral between-population genetic variation was converted into within-population variation, raising the possibility that hybridization between colonizing lineages contributed to the ecological success of sticklebacks in Switzerland.

Prognostic value of microvascular invasion in predicting the cancer specific survival and risk of metastatic disease in renal cell carcinoma: a multicenter investigation

While microvascular invasion is an accepted risk factor in various cancers, its prognostic role in renal cell carcinoma is still unclear. Therefore, a large multicenter study examining the experience of 5 international institutions was performed to evaluate the prognostic value of microvascular invasion in the occurrence of metastases and cancer specific survival.

Island Invasion by a Threatened Tree Species: Evidence for Natural Enemy Release of Mahogany (Swietenia macrophylla) on Dominica, Lesser Antilles

Despite its appeal to explain plant invasions, the enemy release hypothesis (ERH) remains largely unexplored for tropical forest trees. Even scarcer are ERH studies conducted on the same host species at both the community and biogeographical scale, irrespective of the system or plant life form. In Cabrits National Park, Dominica, we observed patterns consistent with enemy release of two introduced, congeneric mahogany species, Swietenia macrophylla and S. mahagoni, planted almost 50 years ago. Swietenia populations at Cabrits have reproduced, with S. macrophylla juveniles established in and out of plantation areas at densities much higher than observed in its native range. Swietenia macrophylla juveniles also experienced significantly lower leaf-level herbivory (~3.0%) than nine co-occurring species native to Dominica (8.4–21.8%), and far lower than conspecific herbivory observed in its native range (11%–43%, on average). These complimentary findings at multiple scales support ERH, and confirm that Swietenia has naturalized at Cabrits. However, Swietenia abundance was positively correlated with native plant diversity at the seedling stage, and only marginally negatively correlated with native plant abundance for stems ≥1-cm dbh. Taken together, these descriptive patterns point to relaxed enemy pressure from specialized enemies, specifically the defoliator Steniscadia poliophaea and the shoot-borer Hypsipyla grandella, as a leading explanation for the enhanced recruitment of Swietenia trees documented at Cabrits.

Glycoprotein D of bovine herpesvirus 5 (BoHV-5) confers an extended host range to BoHV-1 but does not contribute to invasion of the brain

Bovine herpesvirus 1 (BoHV-1) and BoHV-5 are closely related pathogens of cattle, but only BoHV-5 is considered a neuropathogen. We engineered intertypic gD exchange mutants with BoHV-1 and BoHV-5 backbones in order to address their in vitro and in vivo host ranges, with particular interest in invasion of the brain. The new viruses replicated in cell culture with similar dynamics and to titers comparable to those of their wild-type parents. However, gD of BoHV-5 (gD5) was able to interact with a surprisingly broad range of nectins. In vivo, gD5 provided a virulent phenotype to BoHV-1 in AR129 mice, featuring a high incidence of neurological symptoms and early onset of disease. However, only virus with the BoHV-5 backbone, independent of the gD type, was detected in the brain by immunohistology. Thus, gD of BoHV-5 confers an extended cellular host range to BoHV-1 and may be considered a virulence factor but does not contribute to the invasion of the brain.