Kinder- und Jugendhilfereport 2018. Eine kennzahlenbasierte Analyse

Der Kinder- und Jugendhilfereport 2018, die zentrale Publikation der Arbeitsstelle Kinder- und Jugendhilfestatistik (AKJStat), beschreibt umfassend die aktuelle Situation und die neuere Entwicklung der Kinder- und Jugendhilfe. Grundlage sind die Daten der amtlichen Kinder- und Jugendhilfestatistik. Erstmals wird die Kinder- und Jugendhilfe auf der Basis von Kennzahlen dargestellt und analysiert. Der Report ermöglicht einen schnellen und zuverlässigen Überblick über zentrale Arbeitsfelder und wichtige Aufgabengebiete. (DIPF/Orig.

Promotion of Glioblastoma Cell Motility by Enhancer of Zeste Homolog 2 (EZH2) Is Mediated by AXL Receptor Kinase

<div><p>Enhancer of zeste homolog 2 (EZH2) is the catalytic subunit of the Polycomb-repressive complex 2 (PRC2) that epigenetically silences gene transcription through histone H3 lysine trimethylation (H3K27me3). EZH2 has been implicated in stem cell maintenance and is overexpressed in hematological and solid malignancie`s including malignant glioma. EZH2 is thought to promote tumor progression by silencing tumor suppressor genes. Hence pharmacological disruption of the PRC2 is an attractive therapeutic strategy for cancer treatment. Here we show that EZH2 is expressed in human glioma and correlates with malignancy. Silencing of EZH2 reduced glioma cell proliferation and invasiveness. While we did not observe induction of cell cycle-associated tumor suppressor genes by silencing or pharmacological inhibition of EZH2, microarray analyses demonstrated a strong transcriptional reduction of the AXL receptor kinase. Neither histone nor DNA methylation appeared to be involved in the positive regulation of AXL by EZH2. Silencing AXL mimicked the antiinvasive effects of EZH2 knockdown. Finally, AXL expression is found in human gliomas with high EZH2 expression. Collectively these data suggest that EZH2 drives glioma invasiveness via transcriptional control of AXL independent of histone or DNA methylation.</p> </div

EZH2-knockdown inhibits proliferation and invasion of human malignant glioma cells.

<p><b>A </b><i>EZH2</i> transcript expression was decreased 24 h after si<i>EZH2</i> treatment (left). Western blot showing EZH2 protein expression, 120 h after knockdown by siRNA (right). Tubulin served as loading control. <b>B</b> Cell cycle analysis of U87MG glioma cells 120 h after specific knockdown of <i>EZH2</i> (si<i>EZH2</i>, right) or scrambled control (siC, left). <b>C</b> Invasion of U87MG glioma cells with transient <i>EZH2</i> knockdown (lower panel, black bar) through a matrigel-coated boyden chamber in comparison to control (upper panel, white bar). <b>D</b> Representative dot plots and corresponding analysis of Annexin-V-FITC/DAPI co-staining of U87MG glioma cells untreated or treated for 120 h with 10 µM DZNep. The lower left quadrants represent the living cells (low Annexin-V-FITC-/DAPI-signal), the lower right quadrants represents early apoptosis (low DAPI- and strong Annexin-V-FITC-signal) and the upper right late apoptotic/necrotic cells (double-stained cells). <b>E</b> H3K27me3 methylation was strongly decreased in whole cell lysates of U87MG glioma cells after treatment with 5 µM DZNep or after specific knockdown of <i>EZH2</i> for 120 h. Tubulin served as loading control. <b>F</b> Cell cycle analysis of U87MG glioma cells untreated or treated for 120 h with 500 nM and 5 µM DZNep. <b>G</b> Analysis of nestin expression in S24 glioma-initiating cells untreated (left) or treated with 5 µM DZNep for 120 h (right) by flow cytometry. <b>H</b> Matrigel boyden chamber assay of U87MG glioma cells untreated (upper panel, white bar) and treated (lower panel, black bar) with 5 µM DZNep. Asterisk indicates * (p<0.05). Error bars indicate s.e.m.</p

Methylation independent regulation of AXL.

<p><b>A </b><i>AXL</i> mRNA expression of U87MG human glioma cells treated with 5 µM DZNep for 120 h in comparison to control. <b>B</b> Analysis of AXL protein expression in U87MG glioma cells untreated (solid line) or treated with 5 µM DZNep for 120 h (dashed line) stained with an AXL specific antibody (blue) or an isotype control antibody (red). <b>C</b> Comparison of genes with decreased (upper Venn-diagram) or increased (lower Venn diagram) expression after 120 h of <i>EZH2</i> knockdown (purple) or DZNep treatment (blue) of U87MG glioma cells. Cutoff: 1.5 fold change. <b>D </b><i>AXL</i> mRNA expression of U87MG human malignant glioma cells after 96 h treatment with the DNA methylation inhibitor 5-aza-2′-deoxycytidine (5-aza) (black bars) or DMSO (white bar). <b>E </b><i>EZH2</i> and <i>AXL</i> mRNA expression of U87MG glioma cells after the stimulation with indicated concentrations of the histone deacetylase inhibitors suberoylanilide hydroxamic acid (SAHA) or trichostatin A (TSA) (black bars) or DMSO (white bars) for 24 h. Asterisk indicates * (p<0.05). Error bars indicate s.e.m.</p

Transcriptional profiling of EZH2-knockdown in human malignant glioma cells.

<p><b>A</b> mRNA expression of various known EZH2 target genes 120 h after siRNA mediated <i>EZH2</i> knockdown in U87MG glioma cells. <b>B</b> Table of known EZH2 target genes, which were regulated as indicated in response to <i>EZH2</i> knockdown compared to control in U87MG glioma cells. <b>C</b> Heatmap of the ten most regulated genes in response to specific knockdown of EZH2 by siRNA after 120 h in U87MG cells. Upregulated genes are indicated in blue, downregulated genes in yellow. <b>D</b> AXL protein expression in the human glioma cell line U87MG (left) and the human GIC S24 (right) by flow cytometry. The cells were stained with an AXL specific antibody: blue or an isotype control: red. <b>E </b><i>AXL</i> mRNA expression levels in U87MG human glioma cells incubated for 120 h with scrambled control siRNA (siC) or <i>EZH2</i> specific siRNA (si<i>EZH2</i>). <b>F</b> Analysis of AXL protein expression in U87MG glioma cells treated with siC (solid line) or si<i>EZH2</i> (dashed line) for 120 h stained with a AXL specific antibody: blue, isotype control: red. Asterisk indicates * (p<0.05). Error bars indicate s.e.m. NS: not significant.</p

EZH2 expression in human malignant glioma.

<p><b>A</b> EZH2 expression (red) in human glioma sections of different WHO grades: astrocytoma grade II (top), astrocytoma grade III (middle), glioblastoma (bottom). Magnification 200x, inset 400x. <b>B</b> Box-plot of EZH2 expression in human brain tumors of increasing malignancy (WHO grade II: n = 10, mean = 6, SD = 18.97; WHO grade III: n = 15, mean = 40.67, SD = 56.91; GBM: n = 12, mean = 135, SD = 98.59; r² = 0.385, P = 0.024). <b>C</b> EZH2 expression (red) in glioblastoma sections in close proximity to necrotic areas. Arrows indicate the necrotic area. Magnification 100x. <b>D </b><i>EZH2</i> mRNA expression in A172, LN18, U87MG human malignant glioma cells, S24, T269 human glioma-initiating cells (GIC), human astrocytes and human mesenchymal stem cells (MSC) measured by qRT-PCR. <b>E</b> EZH2 protein expression in A172, LN18, U87MG human malignant glioma cells, S24, T269 human GIC, human astrocytes and human MSC. Tubulin served as loading control. Asterisk indicates * (p<0.05). Error bars indicate s.e.m.</p