Genomic and Expression Profiling of Glioblastoma Stem Cell-Like Spheroid Cultures Identifies Novel Tumor-Relevant Genes Associated with Survival
Genomic and Expression Profiling of Glioblastoma Stem Cell-Like Spheroid Cultures Identifies Novel Tumor-Relevant Genes Associated with Survival
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DOI：
10.1158/1078-0432.ccr-09-0695
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发表时间：
2009-11-01
影响因子：
11.5
通讯作者：
Radlwimmer, Bernhard
中科院分区：
文献类型：
--
作者：
Ernst, Aurelie;Hofmann, Stefanie;Radlwimmer, Bernhard
目的：胶质母细胞瘤球状体培养物富含肿瘤干细胞样细胞，因此可能比常规单层培养物更能代表相应的原发性肿瘤。我们利用神经胶质瘤球体培养模型，找到新的肿瘤相关genes.Experimental设计：我们进行了基于阵列的比较基因组杂交的球体文化来自20胶质母细胞瘤。应用基于微阵列的基因表达分析来确定与正常脑组织和胶质瘤球体培养物中的非肿瘤性脑球体相比具有差异表达的基因。通过组织芯片上的免疫组织化学测定三种候选者的蛋白表达水平，并与临床结果相关联。PDPN的功能分析done.Results：基因组的变化在球体文化密切类似于在原发性肿瘤中检测到相应的患者。相反，从相同患者建立的血清生长单层培养物中的基因组变化与相应的原发性肿瘤不匹配。基于微阵列的胶质母细胞瘤球状体培养物的基因表达分析确定了一组新的候选基因相对于正常脑被上调或下调。对20例胶质母细胞瘤临床样本中8个候选基因进行实时定量PCR分析，验证了微阵列结果。组织芯片的免疫组化显示AJAP 1、EMP 3和PDPN的表达与星形胶质细胞胶质瘤患者的总生存率显著相关。侵袭能力和RhoA活性下降PDPN沉默spheroids.Conclusion：我们确定了一组新的候选基因，可能发挥作用，在胶质母细胞瘤的发病机制和牵连AJAP 1，EMP 3，PDPN作为分子标记物与胶质瘤患者的临床结果。(Clin癌症研究2009;15（21）：6541-50）
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)