Characterisation of differential gene expression of soft tissue sarcomas by microarray technology
Characterisation of differential gene expression of soft tissue sarcomas by microarray technology
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微阵列技术表征软组织肉瘤差异基因表达
DOI：
10.1038/14314
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发表时间：
1999
期刊：
影响因子：
30.8
通讯作者：
I. Andrulis
中科院分区：
文献类型：
--
作者：
N. Gokgoz;J. Wunder;I. Andrulis
阵列技术为从临床样本中鉴定诊断和预后标志物以及药物靶点创造了机会。我们已经开发了一种中等密度的斑点寡核苷酸阵列平台，用于监测人类肿瘤中的基因表达和基因组扩增和丢失事件。点样寡核苷酸阵列是灵敏的，稳健的，并允许快速掺入新的序列信息。我们还设计了一个单一的定制Affyscore GeneChip™阵列，通过经验选择探针，在广泛的样品中保持一致的性能，监测35，000个基因和EST。整合来自Affytelum和Eos寡核苷酸阵列的基因表达数据，结合基因组扩增数据，已经鉴定了乳腺癌中具有增加的mRNA表达和基因组代表性的许多转录物。我们进一步生成了一个正常组织图谱数据库，使我们能够筛选潜在的药物线索，以发现可能的毒副作用。然后，通过DNA微阵列鉴定的候选疾病基因被用于通过原位杂交以高度平行的方式询问排列的存档肿瘤组织。这些肿瘤组织微阵列允许以单细胞分辨率快速表征表达。此外，基因表达可能与数百个具有已知临床结果的存档肿瘤的肿瘤分级和分期相关。使用这些多重阵列技术，我们已经鉴定了许多预期编码肿瘤特异性蛋白的转录物，这些蛋白是乳腺癌治疗干预、诊断和预后的潜在靶点。
Array technologies create an opportunity to identify diagnostic and prognostic markers, as well as drug targets, from clinical samples. We have developed a middensity spotted oligonucleotide array platform for monitoring gene expression and genomic amplification and loss events in human tumours. The spotted oligo arrays are sensitive, robust and allow rapid incorporation of new sequence information. We have also designed a single custom Affymetrix GeneChip™ array to monitor 35,000 genes and ESTs by empirically choosing probes on the basis of consistent performance over a wide range of samples. Integration of gene expression data from Affymetrix and Eos oligo arrays, in conjunction with genome amplification data, has identified a number of transcripts with increased mRNA expression and genomic representation in breast cancer. We have further generated a normal tissue atlas database that allows us to screen potential drug leads for possible toxic side effects. Candidate disease genes identified by DNA microarrays are then used to interrogate arrayed archival tumour tissues in a highly parallel fashion by in situ hybridisation. These tumour tissue microarrays allow for the rapid characterisation of expression at single cell resolution. In addition, gene expression can be associated with tumour grade and stage on hundreds of archival tumours with known clinical outcomes. Using these multiple array technologies we have identified a number of transcripts expected to encode tumour-specific proteins which are potential targets for therapeutic intervention, diagnosis and prognosis of breast cancer.