Inhibition of the tyrosine phosphatase SHP-2 suppresses angiogenesis in vitro and in vivo

Endothelial cell survival is indispensable to maintain endothelial integrity and initiate new vessel formation. We investigated the role of SHP-2 in endothelial cell survival and angiogenesis in vitro as well as in vivo. SHP-2 function in cultured human umbilical vein and human dermal microvascular endothelial cells was inhibited by either silencing the protein expression with antisense-oligodesoxynucleotides or treatment with a pharmacological inhibitor (PtpI IV). SHP-2 inhibition impaired capillary-like structure formation (p < 0.01; n = 8) in vitro as well as new vessel growth ex vivo (p < 0.05; n = 10) and in vivo in the chicken chorioallantoic membrane (p < 0.01, n = 4). Additionally, SHP-2 knock-down abrogated fibroblast growth factor 2 (FGF-2)-dependent endothelial proliferation measured by MTT reduction ( p ! 0.01; n = 12). The inhibitory effect of SHP-2 knock-down on vessel growth was mediated by increased endothelial apoptosis ( annexin V staining, p ! 0.05, n = 9), which was associated with reduced FGF-2-induced phosphorylation of phosphatidylinositol 3-kinase (PI3-K), Akt and extracellular regulated kinase 1/2 (ERK1/2) and involved diminished ERK1/2 phosphorylation after PI3-K inhibition (n=3). These results suggest that SHP-2 regulates endothelial cell survival through PI3-K-Akt and mitogen-activated protein kinase pathways thereby strongly affecting new vessel formation. Thus, SHP-2 exhibits a pivotal role in angiogenesis and may represent an interesting target for therapeutic approaches controlling vessel growth. Copyright (C) 2007 S. Karger AG, Basel

Human RNA Polymerase II-Association Factor 1 (hPaf1/PD2) Regulates Histone Methylation and Chromatin Remodeling in Pancreatic Cancer

Change in gene expression associated with pancreatic cancer could be attributed to the variation in histone posttranslational modifications leading to subsequent remodeling of the chromatin template during transcription. However, the interconnected network of molecules involved in regulating such processes remains elusive. hPaf1/PD2, a subunit of the human PAF-complex, involved in the regulation of transcriptional elongation has oncogenic potential. Our study explores the possibility that regulation of histone methylation by hPaf1 can contribute towards alteration in gene expression by nucleosomal rearrangement. Here, we show that knockdown of hPaf1/PD2 leads to decreased di- and tri-methylation at histone H3 lysine 4 residues in pancreatic cancer cells. Interestingly, hPaf1/PD2 colocalizes with MLL1 (Mixed Lineage Leukemia 1), a histone methyltransferase that methylates H3K4 residues. Also, a reduction in hPaf1 level resulted in reduced MLL1 expression and a corresponding decrease in the level of CHD1 (Chromohelicase DNA-binding protein 1), an ATPase dependent chromatin remodeling enzyme that specifically binds to H3K4 di and trimethyl marks. hPaf1/PD2 was also found to interact and colocalize with CHD1 in both cytoplasmic and nuclear extracts of pancreatic cancer cells. Further, reduced level of CHD1 localization in the nucleus in hPaf1/PD2 Knockdown cells could be rescued by ectopic expression of hPaf1/PD2. Micrococcal nuclease digestion showed an altered chromatin structure in hPaf1/PD2-KD cells. Overall, our results suggest that hPaf1/PD2 in association with MLL1 regulates methylation of H3K4 residues, as well as interacts and regulates nuclear shuttling of chromatin remodeling protein CHD1, facilitating its function in pancreatic cancer cells

Downregulated parafibromin expression is a promising marker for pathogenesis, invasion, metastasis and prognosis of gastric carcinomas

Parafibromin is a protein encoded by the hyperparathyroidism 2 oncosuppressor gene and its downregulated expression is involved in pathogenesis of parathyroid carcinomas. To clarify the roles of parafibromin expression in tumourigenesis and progression of gastric carcinomas, it was examined by immunohistochemistry (IHC) on tissue microarray containing gastric carcinomas (n = 508), adenomas (n = 45) and gastritis (n = 49) with a comparison of its expression with clinicopathological parametres of carcinomas. Gastric carcinoma cell lines (MKN28, AGS, MKN45, KATO-III and HGC-27) were studied for parafibromin expression by IHC and western blot. Parafibromin expression was localised in the nucleus of gastric epithelial cells, adenoma, carcinoma cells and cell lines. Its expression was gradually decreased from gastritis to gastric carcinoma, through gastric adenomas (p < 0.05) and inversely correlated with tumour size, depth of invasion, lymphatic invasion, lymph node metastasis and Union Internationale Contre le Cancer (UICC) staging (p < 0.05) but not with sex or venous invasion (p > 0.05). Parafibromin was strongly expressed in older carcinoma patients compared with younger ones (p < 0.05). There was stronger positivity of parafibromin in intestinal-type than diffuse-type carcinomas (p < 0.05). Univariate analysis indicated cumulative survival rate of patients with positive parafibromin expression to be higher than without its expression (p < 0.05). Multivariate analysis showed that age, tumour size, depth of invasion, lymphatic invasion, lymph node metastasis, UICC staging and Lauren’s classification but not sex, venous invasion or parafibromin expression were independent prognostic factors for carcinomas(p < 0.05). Downregulated parafibromin expression possibly contributed to pathogenesis, growth, invasion and metastasis of gastric carcinomas. It was considered as a promising marker to indicate the aggressive behaviours and prognosis of gastric carcinomas

The EIF4EBP3 translational repressor is a marker of CDC73 tumor suppressor haploinsufficiency in a parathyroid cancer syndrome

Germline mutation of the tumor suppressor gene CDC73 confers susceptibility to the hyperparathyroidism-jaw tumor syndrome associated with a high risk of parathyroid malignancy. Inactivating CDC73 mutations have also been implicated in sporadic parathyroid cancer, but are rare in sporadic benign parathyroid tumors. The molecular pathways that distinguish malignant from benign parathyroid transformation remain elusive. We previously showed that a hypomorphic allele of hyrax (hyx), the Drosophila homolog of CDC73, rescues the loss-of-ventral-eye phenotype of lobe, encoding the fly homolog of Akt1s1/ PRAS40. We report now an interaction between hyx and Tor, a central regulator of cell growth and autophagy, and show that eukaryotic translation initiation factor 4E-binding protein (EIF4EBP), a translational repressor and effector of mammalian target of rapamycin (mTOR), is a conserved target of hyx/CDC73. Flies heterozygous for Tor and hyx, but not Mnn1, the homolog of the multiple endocrine neoplasia type 1 (MEN1) tumor suppressor associated with benign parathyroid tumors, are starvation resistant with reduced basal levels of Thor/4E-BP. Human peripheral blood cell levels of EIF4EBP3 were reduced in patients with CDC73, but not MEN1, heterozygosity. Chromatin immunoprecipitation demonstrated occupancy of EIF4EBP3 by endogenous parafibromin. These results show that EIF4EBP3 is a peripheral marker of CDC73 function distinct from MEN1-regulated pathways, and suggest a model whereby starvation resistance and/or translational de-repression contributes to parathyroid malignant transformation

Preferenda thermiques des champignons associés à lps sexdentatus Boern. et Tomicus piniperda L. ( Coleoptera : Scolytidae)

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Phosphoinositide 3-kinases in lysophosphatidic acid signaling: regulation and cross-talk with the Ras/mitogen-activated protein kinase pathway

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Observations symptomatologiques et rôles possibles d'Ophiostoma minus Hedgc. (ascomycète : Ophiostomatales) et de Tomicus piniperda L. (Coleoptera : Scolytidae) dans le dépérissement du pin sylvestre en forêt d'Orléans

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Cinétique de croissance des champignons associés à lps sexdentatus Boern et Tomicus piniperda L (Coleoptera : Scolytidae) et des réactions de défense des pins sylvestres (Pinus sylvestris L) inoculés

A 3 saisons différentes de l'année, 5 espèces de champignons associées à I sexdentatus et T piniperda ont été inoculées à des pins sylvestres. Des rondins fraîchement coupés ont aussi été inoculés au laboratoire, à 1 seule saison. Le suivi pendant 60 j de la cinétique de croissance des réactions de défense, estimées par leur longueur ou par la quantité de résine (quantité totale et concentration), ainsi que la cinétique de croissance des champignons à l'intérieur des réactions, ont permis d'avancer diverses hypothèses quant au fonctionnement des relations pin sylvestre-scolytides-champignons. Le rôle potentiel de chacun des champignons dans l'induction des réactions de défense, ou dans la mort des arbres lorsque ceux-ci sont attaqués par les scolytes, est discuté. La réaction aurait pour rôle de stopper ou de fortement ralentir la croissance de ces champignons, en attendant que le bourrelet cicatriciel isole complètement les tissus agressés. La longueur de la réaction dans le phloème serait conditionnée par les capacités de diffusion de l'éliciteur, alors que la quantité de résine accumulée dépendrait de la quantité d'éliciteur. L'arrêt de l'accumulation de la résine en un lieu donné ne serait pas dû à la saturation des tissus, mais à un arrêt de l'élicitation en ce lieu. Après la fin de la croissance de la réaction, il y aurait homogénéisation de la répartition de la résine à l'intérieur des tissus concernés, par diffusion de la résine des zones à concentration élevée vers les zones à concentration plus faible et, dans certains cas, par continuation de la synthèse résinique. Il est aussi proposé de définir 3 paramètres représentatifs de la réaction : rapidité, intensité, efficacité.Kinetics of growth of fungi associated with lps sexdentatus and Tomicus piniperda (Coleoptera: Scolytidae) and of the defence reaction of the inoculated scots pines. At 3 different seasons in the year, 5 species of fungi associated with I sexdentatus and T piniperda were inoculated into scots pine trees. Logs were also inoculated once during the year in the lab. The reactions were estimated by their length and by the quantity of resin (both total and concentration). The percentage of sieve cells filled with resin was appreciated by histological sections made in each reaction, at 10 and 30 mm from the point of inoculation and in the middle between this point and the end of the reaction. The kinetics of growth of the fungi and of the defence reactions (length, resin, histology) were followed for 60 days. The temperature and defence reactions were not sufficient to explain the differences in the kinetics of growth between fungi and between seasons (figs 1 and 7). Factors related to intrinsic characteristics of the fungi and to physiological state of the tree may play a role. The possible role of each fungus is discussed according to the induction of the reaction and to tree death when attacked by bark beetle populations. The wound periderm does not seem to be directly responsible for stopping the fungal growth. In return, the considerable slowing down of this growth, followed by a slowing down of the development of the reaction length (fig 3) may give sufficient time for the wound periderm to be built, to insulate the wounded tissues completely and to contain the aggressors. The parallel between the kinetics of growth of the fungi and the kinetics of resin accumulation suggests that the quantity of resin would depend upon the quantity of elicitor produced during the fungal growth. The reaction length in the phloem would depend on the diffusion ability of the elicitor. The end of the resin accumulation at a given place would not depend upon tissue saturation, but rather on the end of the elicitation at this place (compare figs 3, 5 and table III). After the end of the reaction growth, an homogenization process of the resin distribution may take place inside the reaction, by a diffusion of the resin from the highly concentrated zones to the less concentrated zones and, in some cases, by a continuation of the resin synthesis. A definition of 3 parameters representative for the defence reaction is proposed: speed, intensity, efficiency