Genomic and Expression Profiling of Glioblastoma Stem Cell-Like Spheroid Cultures Identifies Novel Tumor-Relevant Genes Associated with Survival

被引：146

作者：

Ernst, Aurelie

Hofmann, Stefanie ^{[4
]}

Ahmadi, Rezvan ^{[5
]}

Becker, Natalia

Korshunov, Andrey ^{[2
,6
]}

Engel, Felix

Hartmann, Christian ^{[2
,6
]}

Felsberg, Joerg ^{[7
]}

Sabe, Michael ^{[8
]}

Peterziel, Heike ^{[3
]}

Durchdewald, Moritz ^{[3
]}

Hess, Jochen ^{[3
]}

Barbus, Sebastian

Campos, Benito ^{[5
]}

Starzinski-Powitz, Anna ^{[9
]}

Unterberg, Andreas ^{[5
]}

Reifenberger, Guido ^{[7
]}

Lichter, Peter

Herold-Mende, Christel ^{[5
]}

Radlwimmer, Bernhard ^{[1
]}

机构：

[1] Univ Heidelberg, German Canc Res Ctr, Div Mol Genet, D-69120 Heidelberg, Germany

[2] Univ Heidelberg, Clin Cooperat Unit Neuropathol, D-69120 Heidelberg, Germany

[3] Univ Heidelberg, German Canc Res Ctr, Div Transduct & Growth Control, D-69120 Heidelberg, Germany

[4] Univ Heidelberg, Natl Ctr Tumor Dis, Div Gastrooncol, D-69120 Heidelberg, Germany

[5] Univ Heidelberg, Dept Neurosurg, Div Neurosurg Res, D-69120 Heidelberg, Germany

[6] Univ Heidelberg, Dept Neuropathol, D-69120 Heidelberg, Germany

[7] Univ Dusseldorf, Dept Neuropathol, Dusseldorf, Germany

[8] Univ Dusseldorf, Dept Neurosurg, Dusseldorf, Germany

[9] Goethe Univ Frankfurt, Inst Cell Biol & Neurosci, Ctr Membrane Prote, Frankfurt, Germany

来源：

CLINICAL CANCER RESEARCH | 2009年 / 15卷 / 21期

关键词：

MICROARRAY ANALYSIS; MALIGNANT GLIOMAS; CANCER; LINES; PROGRESSION; GENERATION; SUPPRESSOR; CARCINOMA; REVEALS; 19Q13.3;

D O I：

10.1158/1078-0432.CCR-09-0695

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

Purpose: Glioblastoma spheroid cultures are enriched in tumor stem-like cells and therefore may be more representative of the respective primary tumors than conventional monolayer cultures. We exploited the glioma spheroid culture model to find novel tumor-relevant genes. Experimental Design: We carried out array-based comparative genomic hybridization of spheroid cultures derived from 20 glioblastomas. Microarray-based gene expression analysis was applied to determine genes with differential expression compared with normal brain tissue and to nonneoplastic brain spheroids in glioma spheroid cultures. The protein expression levels of three candidates were determined by immunohistochemistry on tissue microarrays and correlated with clinical outcome. Functional analysis of PDPN was done. Results: Genomic changes in spheroid cultures closely resembled those detected in primary tumors of the corresponding patients. In contrast, genomic changes in serum-grown monolayer cultures established from the same patients did not match well with the respective primary tumors. Microarray-based gene expression analysis of glioblastoma spheroid cultures identified a set of novel candidate genes being upregulated or downregulated relative to normal brain. Quantitative real-time PCR analyses of 8 selected candidate genes in 20 clinical glioblastoma samples validated the microarray findings. Immunohistochemistry on tissue microarrays revealed that expression of AJAP1, EMP3, and PDPN was significantly associated with overall survival of astrocytic glioma patients. Invasive capacity and RhoA activity were decreased in PDPN-silenced spheroids. Conclusion: We identified a set of novel candidate genes that likely play a role in glioblastoma pathogenesis and implicate AJAP1, EMP3, and PDPN as molecular markers associated with the clinical outcome of glioma patients. (Clin Cancer Res 2009;15(21):6541-50)

引用

页码：6541 / 6550

页数：10

共 37 条

[1] EMP3, a myelin-related gene located in the critical 19q13.3 region, is epigenetically silenced and exhibits features of a candidate tumor suppressor in glioma and neuroblastoma [J].