Galamb Orsolya, Sipos Ferenc, Spisák Sándor, Galamb Barnabás, Krenács Tibor, Valcz Gábor, Tulassay Zsolt, Molnár Béla
2nd Department of Medicine, Semmelweis University, Szentkiralyi str. 46, Budapest, Hungary.
Cell Oncol. 2009;31(1):19-29. doi: 10.3233/clo-2009-0458.
As most colorectal cancers (CRC) develop from villous adenomas, studying alterations in gene expression profiles across the colorectal adenoma-dysplasia-carcinoma sequence may yield potential biomarkers of disease progression.
Total RNA was extracted, amplified, and biotinylated from colonic biopsies of 15 patients with CRC, 15 with villous adenoma and 8 normal controls. Gene expression profiles were evaluated using HGU133Plus2.0 microarrays and disease progression associated data were validated with RT-PCR. The potential biomarkers were also tested at the protein level using tissue microarray samples of 103 independent and 16 overlapping patients.
17 genes were validated to show sequentially altered expression at mRNA level through the normal-adenoma-dysplasia-carcinoma progression. Prostaglandin-D2 receptor (PTGDR) and amnionless homolog (AMN) genes revealed gradually decreasing expression while the rest of 15 genes including osteonectin, osteopontin, collagen IV-alpha 1, biglycan, matrix GLAprotein, and von Willebrand factor demonstrated progressively increasing expression. Similar trends of expression were confirmed at protein level for PTGDR, AMN, osteopontin and osteonectin.
Downregulated AMN and PTGDR and upregulated osteopontin and osteonectin were found as potential biomarkers of colorectal carcinogenesis and disease progression to be utilized for prospective biopsy screening both at mRNA and protein levels. Gene alterations identified here may also add to our understanding of CRC progression.
由于大多数结直肠癌（CRC）由绒毛状腺瘤发展而来，研究结直肠腺瘤 - 发育异常 - 癌序列中基因表达谱的变化可能会产生疾病进展的潜在生物标志物。
从15例CRC患者、15例绒毛状腺瘤患者和8例正常对照的结肠活检组织中提取、扩增并生物素化总RNA。使用HGU133Plus2.0微阵列评估基因表达谱，并通过RT-PCR验证与疾病进展相关的数据。还使用103例独立患者和16例重叠患者的组织微阵列样本在蛋白质水平上测试潜在的生物标志物。
验证了17个基因在从正常 - 腺瘤 - 发育异常 - 癌的进展过程中在mRNA水平上显示出顺序改变的表达。前列腺素 - D2受体（PTGDR）和无羊膜同源物（AMN）基因表达逐渐降低，而其余15个基因，包括骨连接蛋白、骨桥蛋白、IV型胶原α1、双糖链蛋白聚糖（双糖链蛋白聚糖）、基质GLA蛋白和血管性血友病因子，表达逐渐增加。PTGDR、AMN、骨桥蛋白和骨连接蛋白在蛋白质水平上也证实了类似的表达趋势。
发现AMN和PTGDR下调以及骨桥蛋白和骨连接蛋白上调是结直肠癌发生和疾病进展的潜在生物标志物，可用于mRNA和蛋白质水平的前瞻性活检筛查。此处鉴定的基因改变也可能增进我们对CRC进展的理解。
