Differential expression of microRNA species in human gastric cancer versus non-tumorous tissues
Differential expression of microRNA species in human gastric cancer versus non-tumorous tissues
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DOI：
10.1111/j.1440-1746.2008.05666.x
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发表时间：
2009-04-01
影响因子：
4.1
通讯作者：
Wang, Yanjun
中科院分区：
文献类型：
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作者：
Guo, Junming;Miao, Ying;Wang, Yanjun
微小RNA（miRNA）在致癌过程中发挥着重要作用。胃癌的总体miRNA表达谱尚未见报道。本研究从胃癌组织和癌旁组织中提取总RNA，将小分子RNA（< 300 nt）进行3 '端延伸，加入poly（A）尾，以确定胃癌组织中miRNA的表达谱。在μ Paraflo（TM）微流控芯片（LC Sciences，Houston，TX，USA）上进行杂交。在通过使用标签特异性Cy 3和Cy 5染料的荧光标记进行杂交检测后，使用激光扫描仪收集杂交图像，并使用Array-Pro图像分析软件（Media Cybernetics，银Spring，MD，USA）进行数字化。为验证上述结果并探讨差异表达的miRNAs的生物学意义，采用免疫组织化学方法检测了在非肿瘤组织中高表达的miRNAs，其中miR-768- 3 p、miR-139- 5 p、miR-378、miR-31、miR-195、miR-497和miR-133 b的表达量最高。其中miR-139- 5 p、miR-497和miR-768- 3 p是首次在非肿瘤组织中发现的。胃癌组织中高表达的miRNAs为miR-20 b、miR-20 a、miR-17、miR-106 a、miR-18 a、miR-21、miR-106 b、miR-18 b、miR-421、miR-340*、miR-19 a和miR-658。其中，miR-340*、miR-421和miR-658首次在癌细胞中发现高表达。发现某些靶基因（如Rb和PTEN）在胃癌组织中的表达降低，据我们所知，这是首次报道这些与胃癌相关的miRNAs。这些新的信息可能提示这些miRNAs在胃癌诊断中的潜在作用。
MicroRNAs (miRNAs) play important roles in carcinogenesis. The global miRNA expression profile of gastric cancer has not been reported. The purpose of the present study was to determine the miRNA expression profile of gastric cancer.Total RNA were first extracted from primary gastric cancer tissues and adjacent non-tumorous tissues and then small isolated RNAs (< 300 nt) were 3'-extended with a poly(A) tail. Hybridization was carried out on a mu Paraflo (TM) microfluidic chip (LC Sciences, Houston, TX, USA). After hybridization detection by fluorescence labeling using tag-specific Cy3 and Cy5 dyes, hybridization images were collected using a laser scanner and digitized using Array-Pro image analysis software (Media Cybernetics, Silver Spring, MD, USA). To validate the results and investigate the biological meaning of differential expressed miRNAs, immunohistochemistry was used to detect the differential expression of target genes.The most highly expressed miRNAs in non-tumorous tissues were miR-768-3p, miR-139-5p, miR-378, miR-31, miR-195, miR-497 and miR-133b. Three of them, miR-139-5p, miR-497 and miR-768-3p, were first found in non-tumorous tissues. The most highly expressed miRNAs in gastric cancer tissues were miR-20b, miR-20a, miR-17, miR-106a, miR-18a, miR-21, miR-106b, miR-18b, miR-421, miR-340*, miR-19a and miR-658. Among them, miR-340*, miR-421 and miR-658 were first found highly expressed in cancer cells. The expression of some target genes (such as Rb and PTEN) in cancer tissues was found to be decreased.To our knowledge, this is the first report about these miRNAs associated with gastric cancer. This new information may suggest the potential roles of these miRNAs in the diagnosis of gastric cancer.