mRNA expression, functional profiling and multivariate classification of colon biopsy specimen by cDNA overall glass microarray
mRNA expression, functional profiling and multivariate classification of colon biopsy specimen by cDNA overall glass microarray
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DOI：
10.3748/wjg.v12.i43.6998
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发表时间：
2006-11-21
影响因子：
4.3
通讯作者：
Molnar, Bela
中科院分区：
文献类型：
--
作者：
Galamb, Orsolya;Sipos, Ferenc;Molnar, Bela
目的：目的：了解结肠活检组织局部病理生理学改变和基于基因本体的炎症性和肿瘤性疾病的功能分类。方法：从冰冻活检组织中提取总RNA，T7-法扩增。通过Atlas Glass 1 K微阵列评估表达谱。在微阵列质量控制后，可从10个腺瘤、6个结直肠腺癌（CRC）和6个炎性肠病（IBD）获得适用的数据。进行多变量统计和细胞功能分析。结果：采用热休克转录因子-1、bystin样蛋白、钙颗粒蛋白A、TRAIL受体3 4个参数对22例标本进行判别分析，结果正确。IBD样本的特征在于过度表达的趋化因子（C-X-C基序）配体13、复制蛋白A1、E74样因子2和下调的TNF受体相关因子6、BCL 2相互作用的杀伤基因。肿瘤组织中TNF受体相关因子6、复制蛋白A1、E74样因子2表达上调，BCL 2相关X蛋白、calgranulin-A基因表达下调，大肠癌组织中表皮生长因子受体、拓扑异构酶-1、v-jun、TNF受体相关因子6、TRAIL受体3表达上调，RAD 51、RAD 52 DNA修复基因表达下调。蛋白磷酸酶-2A和BCL 2相互作用的杀伤mRNA水平。表皮生长因子受体RT-PCR和免疫组化、拓扑异构酶-1 RT-PCR证实了芯片结果。结论：不同的组织学改变可以通过功能性、多变量分析进行重新分类。病理改变的亚分类需要进一步扩大样本数量的研究。(c)2006年，WIG出版社。All rights reserved.
AIM: To understand the local pathophysiological alterations and gene ontology-based functional classification of colonic biopsies into inflammatory and neoplastic diseases.METHODS: Total RNA was extracted from frozen biopsies and amplified by T7-method. Expression profile was evaluated by Atlas Glass 1K microarrays. After microarray quality control, applicable data were available from 10 adenomas, 6 colorectal adenocarcinomas (CRCs), and 6 inflammatory bowel diseases (IBDs). Multivariate statistical and cell functional analyses were performed. Real-time RT-PCR and immunohistochemistry were used for validation.RESULTS: Discriminant analysis of selected genes, could correctly reclassify all 22 samples using 4 parameters (heat shock transcription factor-1, bystin-like, calgranulin-A, TRAIL receptor 3). IBD samples were characterized by overregulated chemokine (C-X-C motif) ligand 13, replication protein A1, E74-like factor 2 and downregulated TNF receptor-associated factor 6, BCL2-interacting killer genes. In adenomas upregulation of TNF receptor associated factor 6, replication protein A1, E74-like factor 2 and underexpression of BCL2-associated X protein, calgranulin-A genes were found. CRC cases had significantly increased epidermal growth factor receptor, topoisomerase-1, v-jun, TNF receptor-associated factor 6 and TRAIL receptor 3, and decreased RAD51 and RAD52 DNA repair gene, protein phosphatase-2A and BCL2-interacting killer mRNA levels. Epidermal growth factor receptor RT-PCR and immunohistochemistry, topoisomerase-1 RT-PCR confirmed the chip results.CONCLUSION: Different histological alterations can be reclassified by functional, multivariate analysis using cDNA microarrays. Further studies with expanded sample number are needed for subclassification of pathological alterations.(c) 2006 The WIG Press. All rights reserved.