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通过全基因组表达谱分析,鉴定与福尔马林固定、石蜡包埋标本中的非上皮性卵巢癌相关的分子标志物。

Molecular markers associated with nonepithelial ovarian cancer in formalin-fixed, paraffin-embedded specimens by genome wide expression profiling.

机构信息

Department of Physiology, Faculty of Medicine, Universiti Kebangsaan Malaysia, Kuala Lumpur, Malaysia.

出版信息

Kaohsiung J Med Sci. 2012 May;28(5):243-50. doi: 10.1016/j.kjms.2011.11.007. Epub 2012 Apr 6.

Abstract

Nonepithelial ovarian cancer (NEOC) is a rare cancer that is often misdiagnosed as other malignant tumors. Research on this cancer using fresh tissues is nearly impossible because of its limited number of samples within a limited time provided. The study is to identify potential genes and their molecular pathways related to NEOC using formalin-fixed paraffin embedded samples. Total RNA was extracted from eight archived NEOCs and seven normal ovaries. The RNA samples with RNA integrity number >2.0, purity >1.7 and cycle count value <28 cycles were hybridized to the Illumina Whole-Genome DASL assay (cDNA-mediated annealing, selection, extension, and ligation). We analyzed the results using the GeneSpring GX11.0 and FlexArray software to determine the differentially expressed genes. Microarray results were validated using an immunohistochemistry method. Statistical analysis identified 804 differentially expressed genes with 443 and 361 genes as overexpressed and underexpressed in cancer, respectively. Consistent findings were documented for the overexpression of eukaryotic translation elongation factor 1 alpha 1, E2F transcription factor 2, and fibroblast growth factor receptor 3, except for the down-regulated gene, early growth response 1 (EGR1). The immunopositivity staining for EGR1 was found in the majority of cancer tissues. This finding suggested that the mRNA level of a transcript did not always match with the protein expression in tissues. The current gene profile can be the platform for further exploration of the molecular mechanism of NEOC.

摘要

非上皮性卵巢癌(NEOC)是一种罕见的癌症,常被误诊为其他恶性肿瘤。由于在有限的时间内提供的样本数量有限,因此使用新鲜组织对这种癌症进行研究几乎是不可能的。本研究旨在使用福尔马林固定石蜡包埋样本鉴定与 NEOC 相关的潜在基因及其分子途径。从 8 个存档的 NEOC 和 7 个正常卵巢中提取总 RNA。选择 RNA 完整性值>2.0、纯度>1.7 和循环计数值<28 个周期的 RNA 样本进行 Illumina 全基因组 DASL 杂交(cDNA 介导的退火、选择、延伸和连接)。我们使用 GeneSpring GX11.0 和 FlexArray 软件分析结果,以确定差异表达基因。使用免疫组织化学方法验证微阵列结果。统计分析确定了 804 个差异表达基因,其中 443 个和 361 个基因在癌症中分别过表达和低表达。除下调基因早期生长反应 1(EGR1)外,真核翻译延伸因子 1α1、E2F 转录因子 2 和成纤维细胞生长因子受体 3 的过表达一致。免疫阳性染色在大多数癌症组织中发现 EGR1。这一发现表明,转录物的 mRNA 水平并不总是与组织中的蛋白质表达相匹配。当前的基因谱可以作为进一步探索 NEOC 分子机制的平台。

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