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mRNA expression analysis and classification of colonic biopsy samples using oligonucleotide cDNA microarray techniques.

机译：使用寡核苷酸cDNA芯片技术对结肠活检样品进行mRNA表达分析和分类。

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摘要

Despite tremendous progress in the past few decades, certain important aspects regarding the diagnosis, therapy, and follow-up of CRC still remain unsolved. The high incidence and not completely known pathogenetic background, origin, molecular biology of CRC necessitate further research of this disease. In my PhD thesis I searched for biomarkers of the development of colorectal carcinoma, and performed gene expression analysis for colorectal disease classification using whole genomic oligonucleotide and cDNA microarray technology and colonic biopsy samples. I have established that the oligonucleotide whole genomic microarray analyses of biopsy samples wholly fulfill the Affymetrix quality requirements, are highly standard and reproducible. I have established that the Taqman Microfluidic Card System which offers an opportunity for the analysis of the expression levels of 96 genes on 10-100 samples, is particularly suitable for high-throughput, quick and cost efficient RT-PCR validation of gene expression changes detected by microarrays. I have shown, that the sequential overexpression of osteopontin and osteonectin mRNAs and proteins significantly correlates with the progression of the colorectal adenoma-dysplasia-carcinoma sequence. I have identified and validated by RT-PCR ten novel tissue markers which show continuously increasing mRNA expression in line with the colorectal adenoma-dysplasia-carcinoma transition. These are the following: tissue inhibitor of metalloproteinases-1 and -3, von Willebrand factor, interleukin 8, melanoma cell adhesion molecule, thrombospondin 2, collagen 4A1, matrix Gla protein, interleukin 1 receptor antagonist and calumenin. I have established that the prostaglandin D2 receptor and the amnionless homolog are novel, validated sequentially downregulated markers of the colorectal adenoma-dysplasia-carcinoma sequence. During my analyses, I have identified the top 27, 13 and 10 genes associated with adenoma, CRC, and IBD, respectively. Using whole genomic microarrays I have also determined the top100 most differentially expressed discriminatory genes which are suitable for molecular-based discrimination of early (Dukes A and B stage) and advanced (Dukes C and D) colorectal cancer.
机译：尽管在过去的几十年中取得了长足的进步，但是关于CRC的诊断，治疗和随访的某些重要方面仍未解决。 CRC的高发病率且尚不完全清楚，其致病背景，起源，分子生物学都需要对该疾病进行进一步的研究。在我的博士学位论文中，我搜索了结直肠癌发展的生物标志物，并使用完整的基因组寡核苷酸和cDNA微阵列技术以及结肠活检样本对结直肠疾病进行了基因表达分析。我已经确定，活检样品的寡核苷酸全基因组微阵列分析完全符合Affymetrix的质量要求，具有很高的标准性和可重复性。我已经确定，塔克曼微流控卡系统为分析10-100个样品中的96个基因的表达水平提供了机会，特别适合于高通量，快速且经济高效的RT-PCR验证检测到的基因表达变化通过微阵列。我已经证明，骨桥蛋白和骨连接蛋白的mRNA和蛋白的顺序过表达与大肠腺瘤-异型增生-癌序列的进展密切相关。我已经通过RT-PCR鉴定并验证了十种新颖的组织标志物，这些标志物显示了与结直肠腺瘤-异型增生-癌变相一致的不断增加的mRNA表达。它们是：金属蛋白酶-1和-3的组织抑制剂，von Willebrand因子，白介素8，黑素瘤细胞粘附分子，血小板反应蛋白2，胶原蛋白4A1，基质Gla蛋白，白介素1受体拮抗剂和钙粘蛋白。我已经确定，前列腺素D2受体和无羊膜的同系物是新颖的，经验证可依次下调大肠腺瘤-异型增生-癌序列的标志物。在分析过程中，我分别确定了与腺瘤，CRC和IBD相关的前27、13和10个基因。使用完整的基因组微阵列，我还确定了前100个最差异表达的歧视性基因，这些基因适用于早期（杜克A和B期）和晚期（杜克C和D）大肠癌的分子识别。

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  • 作者

    Galamb, Orsolya.;

  • 作者单位

    Semmelweis Egyetem (Hungary).;

  • 授予单位 Semmelweis Egyetem (Hungary).;
  • 学科 Oncology.;Molecular biology.
  • 学位 Dr.
  • 年度 2007
  • 页码 114 p.
  • 总页数 114
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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