Children suffering from neurologic cancers undergoing chemotherapy and radiotherapy are at high risk of reduced neurocognitive abilities likely via damage to proliferating neural stem cells (NSC). Therefore, strategies to protect NSCs are needed. We argue that induced cell-cycle arrest/quiescence in NSCs during cancer treatment can represent such a strategy. Here, we show that hyperpolarization-activated cyclic nucleotide-gated (HCN) ion channels are dynamically expressed over the cell cycle in NSCs, depolarize the membrane potential, underlie spontaneous calcium oscillations and are required to maintain NSCs in the actively proliferating pool. Hyperpolarizing pharmacologic inhibition of HCN channels during exposure to ionizing radiation protects NSCs cells in neurogenic brain regions of young mice. In contrast, brain tumor-initiating cells, which also express HCN channels, remain proliferative during HCN inhibition. IMPLICATIONS: Our finding that NSCs can be selectively rescued while cancer cells remain sensitive to the treatment, provide a foundation for reduction of cognitive impairment in children with neurologic cancers.

• MeSH
• hyperpolarizační iontové kanály řízené cyklickými nukleotidy metabolismus   MeSH
• lidé MeSH
• myši MeSH
• nádory farmakoterapie   MeSH
• nervové kmenové buňky metabolismus   MeSH
• proliferace buněk MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• hyperpolarizační iontové kanály řízené cyklickými nukleotidy MeSH

Increased levels of the chemokine CCL2 in cancer patients are associated with poor prognosis. Experimental evidence suggests that CCL2 correlates with inflammatory monocyte recruitment and induction of vascular activation, but the functionality remains open. Here, we show that endothelial Ccr2 facilitates pulmonary metastasis using an endothelial-specific Ccr2-deficient mouse model (Ccr2ecKO). Similar levels of circulating monocytes and equal leukocyte recruitment to metastatic lesions of Ccr2ecKO and Ccr2fl/fl littermates were observed. The absence of endothelial Ccr2 strongly reduced pulmonary metastasis, while the primary tumor growth was unaffected. Despite a comparable cytokine milieu in Ccr2ecKO and Ccr2fl/fl littermates the absence of vascular permeability induction was observed only in Ccr2ecKO mice. CCL2 stimulation of pulmonary endothelial cells resulted in increased phosphorylation of MLC2, endothelial cell retraction, and vascular leakiness that was blocked by an addition of a CCR2 inhibitor. These data demonstrate that endothelial CCR2 expression is required for tumor cell extravasation and pulmonary metastasis. IMPLICATIONS: The findings provide mechanistic insight into how CCL2-CCR2 signaling in endothelial cells promotes their activation through myosin light chain phosphorylation, resulting in endothelial retraction and enhanced tumor cell migration and metastasis.

• MeSH
• chemokin CCL2 metabolismus   MeSH
• endoteliální buňky metabolismus   patologie   MeSH
• kapilární permeabilita MeSH
• karcinom plic Lewisové krevní zásobení   metabolismus   patologie   sekundární   MeSH
• lehké řetězce myosinu metabolismus   MeSH
• metastázy nádorů MeSH
• myši inbrední C57BL MeSH
• myši knockoutované MeSH
• myši MeSH
• nádory plic krevní zásobení   metabolismus   patologie   sekundární   MeSH
• pohyb buněk fyziologie   MeSH
• receptory CCR2 metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• Ccl2 protein, mouse MeSH Prohlížeč
• Ccr2 protein, mouse MeSH Prohlížeč
• chemokin CCL2 MeSH
• lehké řetězce myosinu MeSH
• receptory CCR2 MeSH

The Hippo pathway effector, Yes-associated protein (YAP), is a transcriptional coactivator implicated in cholangiocarcinoma (CCA) pathogenesis. YAP is known to be regulated by a serine/threonine kinase relay module (MST1/2-LATS1/2) culminating in phosphorylation of YAP at Serine 127 and cytoplasmic sequestration. However, YAP also undergoes tyrosine phosphorylation, and the role of tyrosine phosphorylation in YAP regulation remains unclear. Herein, YAP regulation by tyrosine phosphorylation was examined in human and mouse CCA cells, as well as patient-derived xenograft (PDX) models. YAP was phosphorylated on tyrosine 357 (Y357) in CCA cell lines and PDX models. SRC family kinase (SFK) inhibition with dasatinib resulted in loss of YAPY357 phosphorylation, promoted its translocation from the nucleus to the cytoplasm, and reduced YAP target gene expression, including cell lines expressing a LATS1/2-resistant YAP mutant in which all serine residues were mutated to alanine. Consistent with these observations, precluding YAPY357 phosphorylation by site-directed mutagenesis (YAPY357F) excluded YAP from the nucleus. Targeted siRNA experiments identified LCK as the SFK that most potently mediated YAPY357 phosphorylation. Likewise, inducible CRISPR/Cas9-targeted LCK deletion decreased YAPY357 phosphorylation and its nuclear localization. The importance of LCK in CCA biology was demonstrated by clinical observations suggesting LCK expression levels were associated with early tumor recurrence following resection of CCA. Finally, dasatinib displayed therapeutic efficacy in PDX models. Mol Cancer Res; 16(10); 1556-67. ©2018 AACR.

• MeSH
• adaptorové proteiny signální transdukční genetika   MeSH
• buněčné jádro účinky léků   MeSH
• cholangiokarcinom farmakoterapie   genetika   patologie   MeSH
• cytoplazma účinky léků   MeSH
• dasatinib aplikace a dávkování   MeSH
• fosfoproteiny genetika   MeSH
• fosforylace účinky léků   MeSH
• lidé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• proliferace buněk účinky léků   MeSH
• protein-serin-threoninkinasy genetika   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• signální proteiny YAP MeSH
• signální transdukce účinky léků   MeSH
• skupina kinas odvozených od src-genu antagonisté a inhibitory   genetika   MeSH
• transkripční faktory MeSH
• tyrosin genetika   MeSH
• tyrosinkinasa p56(lck), specifická pro lymfocyty genetika   MeSH
• xenogenní modely - testy protinádorové aktivity MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH
• Názvy látek
• adaptorové proteiny signální transdukční MeSH
• dasatinib MeSH
• fosfoproteiny MeSH
• LATS1 protein, human MeSH Prohlížeč
• LCK protein, human MeSH Prohlížeč
• protein-serin-threoninkinasy MeSH
• signální proteiny YAP MeSH
• skupina kinas odvozených od src-genu MeSH
• transkripční faktory MeSH
• tyrosin MeSH
• tyrosinkinasa p56(lck), specifická pro lymfocyty MeSH
• YAP1 protein, human MeSH Prohlížeč

UNLABELLED: Hypermethylated in cancer 1 (HIC1) represents a prototypic tumor suppressor gene frequently inactivated by DNA methylation in many types of solid tumors. The gene encodes a sequence-specific transcriptional repressor controlling expression of several genes involved in cell cycle or stress control. In this study, a Hic1 allele was conditionally deleted, using a Cre/loxP system, to identify genes influenced by the loss of Hic1. One of the transcripts upregulated upon Hic1 ablation is the toll-like receptor 2 (TLR2). Tlr2 expression levels increased in Hic1-deficient mouse embryonic fibroblasts (MEF) and cultured intestinal organoids or in human cells upon HIC1 knockdown. In addition, HIC1 associated with the TLR2 gene regulatory elements, as detected by chromatin immunoprecipitation, indicating that Tlr2 indeed represents a direct Hic1 target. The Tlr2 receptor senses "danger" signals of microbial or endogenous origin to trigger multiple signaling pathways, including NF-κB signaling. Interestingly, Hic1 deficiency promoted NF-κB pathway activity not only in cells stimulated with Tlr2 ligand, but also in cells treated with NF-κB activators that stimulate different surface receptors. In the intestine, Hic1 is mainly expressed in differentiated epithelial cells and its ablation leads to increased Tlr2 production. Finally, in a chemical-induced mouse model of carcinogenesis, Hic1 absence resulted in larger Tlr2-positive colonic tumors that showed increased proportion of proliferating cells. IMPLICATIONS: The tumor-suppressive function of Hic1 in colon is related to its inhibitory action on proproliferative signaling mediated by the Tlr2 receptor present on tumor cells.

• MeSH
• azoxymethan MeSH
• epitelové buňky MeSH
• genový knockdown MeSH
• karcinogeneze metabolismus   MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• myši transgenní MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádorové supresorové proteiny metabolismus   MeSH
• nádory tračníku MeSH
• NF-kappa B metabolismus   MeSH
• proliferace buněk MeSH
• signální transdukce * MeSH
• síran dextranu MeSH
• střeva cytologie   MeSH
• toll-like receptor 2 metabolismus   MeSH
• transkripční faktory Krüppel-like genetika   metabolismus   MeSH
• upregulace MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• azoxymethan MeSH
• Hic1 protein, mouse MeSH Prohlížeč
• nádorové supresorové proteiny MeSH
• NF-kappa B MeSH
• síran dextranu MeSH
• Tlr2 protein, mouse MeSH Prohlížeč
• toll-like receptor 2 MeSH
• transkripční faktory Krüppel-like MeSH

UNLABELLED: Comparing the gene expression profiles of metastatic and nonmetastatic cells has the power to reveal candidate metastasis-associated genes, whose involvement in metastasis can be experimentally tested. In this study, differentially expressed genes were explored in the v-src-transformed metastatic cell line PR9692 and its nonmetastatic subclone PR9692-E9. First, the contribution of homeodomain only protein X (HOPX) in metastasis formation and development was assessed. HOPX-specific knockdown decreased HOPX expression in the nonmetastatic subclone and displayed reduced cell motility in vitro. Critically, HOPX knockdown decreased the in vivo metastatic capacity in a syngeneic animal model system. Genomic analyses identified a cadre of genes affected by HOPX knockdown that intersected significantly with genes previously found to be differentially expressed in metastatic versus nonmetastatic cells. Furthermore, 232 genes were found in both screens with at least a two-fold change in gene expression, and a number of high-confidence targets were validated for differential expression. Importantly, significant changes were demonstrated in the protein expression level of three metastatic-associated genes (NCAM, FOXG1, and ITGA4), and knockdown of one of the identified HOPX-regulated metastatic genes, ITGA4, showed marked inhibition of cell motility and metastasis formation. These data demonstrate that HOPX is a metastasis-associated gene and that its knockdown decreases the metastatic activity of v-src-transformed cells through altered gene expression patterns. IMPLICATIONS: This study provides new mechanistic insight into a HOPX-regulated metastatic dissemination signature.

• MeSH
• buněčný cyklus MeSH
• down regulace MeSH
• experimentální sarkom genetika   patologie   sekundární   MeSH
• forkhead transkripční faktory genetika   metabolismus   MeSH
• genový knockdown MeSH
• geny src MeSH
• homeodoménové proteiny genetika   metabolismus   MeSH
• kur domácí MeSH
• metastázy nádorů genetika   MeSH
• molekuly buněčné adheze nervové genetika   metabolismus   MeSH
• nádorová transformace buněk genetika   MeSH
• nádorové buněčné linie MeSH
• pohyb buněk MeSH
• ptačí proteiny genetika   metabolismus   MeSH
• regulace genové exprese u nádorů MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
• stanovení celkové genové exprese MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• forkhead transkripční faktory MeSH
• homeodoménové proteiny MeSH
• molekuly buněčné adheze nervové MeSH
• ptačí proteiny MeSH

Hematopoietic transcription factors GATA-1 and PU.1 bind each other on DNA to block transcriptional programs of undesired lineage during hematopoietic commitment. Murine erythroleukemia (MEL) cells that coexpress GATA-1 and PU.1 are blocked at the blast stage but respond to molecular removal (downregulation) of PU.1 or addition (upregulation) of GATA-1 by inducing terminal erythroid differentiation. To test whether GATA-1 blocks PU.1 in MEL cells, we have conditionally activated a transgenic PU.1 protein fused with the estrogen receptor ligand-binding domain (PUER), resulting in activation of a myeloid transcriptional program. Gene expression arrays identified components of the PU.1-dependent transcriptome negatively regulated by GATA-1 in MEL cells, including CCAAT/enhancer binding protein alpha (Cebpa) and core-binding factor, beta subunit (Cbfb), which encode two key hematopoietic transcription factors. Inhibition of GATA-1 by small interfering RNA resulted in derepression of PU.1 target genes. Chromatin immunoprecipitation and reporter assays identified PU.1 motif sequences near Cebpa and Cbfb that are co-occupied by PU.1 and GATA-1 in the leukemic blasts. Significant derepression of Cebpa and Cbfb is achieved in MEL cells by either activation of PU.1 or knockdown of GATA-1. Furthermore, transcriptional regulation of these loci by manipulating the levels of PU.1 and GATA-1 involves quantitative increases in a transcriptionally active chromatin mark: acetylation of histone H3K9. Collectively, we show that either activation of PU.1 or inhibition of GATA-1 efficiently reverses the transcriptional block imposed by GATA-1 and leads to the activation of a myeloid transcriptional program directed by PU.1.

• MeSH
• aktivace transkripce genetika   MeSH
• buněčná diferenciace genetika   MeSH
• HeLa buňky MeSH
• histony genetika   metabolismus   MeSH
• leukemie genetika   metabolismus   patofyziologie   MeSH
• lidé MeSH
• malá interferující RNA MeSH
• myeloidní buňky metabolismus   MeSH
• nádorová transformace buněk genetika   metabolismus   MeSH
• protein CBFB genetika   metabolismus   MeSH
• proteiny vázající zesilovač transkripce CCAAT genetika   metabolismus   MeSH
• protoonkogenní proteiny genetika   MeSH
• regulace genové exprese u nádorů genetika   MeSH
• regulační elementy transkripční genetika   MeSH
• rekombinantní fúzní proteiny genetika   metabolismus   MeSH
• represorové proteiny genetika   metabolismus   MeSH
• RNA interference MeSH
• trans-aktivátory genetika   MeSH
• transkripční faktor GATA1 genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH
• Názvy látek
• CBFB protein, human MeSH Prohlížeč
• CEBPA protein, human MeSH Prohlížeč
• Gata1 protein, mouse MeSH Prohlížeč
• histony MeSH
• malá interferující RNA MeSH
• protein CBFB MeSH
• proteiny vázající zesilovač transkripce CCAAT MeSH
• proto-oncogene protein Spi-1 MeSH Prohlížeč
• protoonkogenní proteiny MeSH
• rekombinantní fúzní proteiny MeSH
• represorové proteiny MeSH
• trans-aktivátory MeSH
• transkripční faktor GATA1 MeSH

Tumor cell invasion is the most critical step of metastasis. Determination of the mode of invasion within the particular tumor is critical for effective cancer treatment. Protease-independent amoeboid mode of invasion has been described in carcinoma cells and more recently in sarcoma cells on treatment with protease inhibitors. To analyze invasive behavior, we compared highly metastatic sarcoma cells with parental nonmetastatic cells. The metastatic cells exhibited a functional up-regulation of Rho/ROCK signaling and, similarly to carcinoma cells, an amoeboid mode of invasion. Using confocal and traction force microscopy, we showed that an up-regulation of Rho/ROCK signaling leads to increased cytoskeletal dynamics, myosin light chain localization, and increased tractions at the leading edge of the cells and that all of these contributed to increased cell invasiveness in a three-dimensional collagen matrix. We conclude that cells of mesenchymal origin can use the amoeboid nonmesenchymal mode of invasion as their primary invading mechanism and show the dependence of ROCK-mediated amoeboid mode of invasion on the increased capacity of cells to generate force.

• MeSH
• aktiny metabolismus   MeSH
• buněčná adheze fyziologie   MeSH
• čipová analýza proteinů MeSH
• cytoskelet metabolismus   patologie   MeSH
• faktory depolymerizující aktin metabolismus   MeSH
• fluorescenční mikroskopie MeSH
• fosforylace MeSH
• invazivní růst nádoru MeSH
• kinázy asociované s Rho genetika   metabolismus   MeSH
• kolagen metabolismus   MeSH
• krysa rodu Rattus MeSH
• kultivované buňky MeSH
• lehké řetězce myosinu metabolismus   MeSH
• Lim-kinasy genetika   metabolismus   MeSH
• magnetismus MeSH
• matrixová metaloproteinasa 2 metabolismus   MeSH
• mezenchymální kmenové buňky metabolismus   patologie   MeSH
• pohyb buněk fyziologie   MeSH
• Rho proteiny vázající GTP genetika   metabolismus   MeSH
• sarkom metabolismus   patologie   MeSH
• upregulace MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aktiny MeSH
• faktory depolymerizující aktin MeSH
• kinázy asociované s Rho MeSH
• kolagen MeSH
• lehké řetězce myosinu MeSH
• Lim-kinasy MeSH
• Limk1 protein, rat MeSH Prohlížeč
• matrixová metaloproteinasa 2 MeSH
• myosin light chain I MeSH Prohlížeč
• Rho proteiny vázající GTP MeSH