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人类胃癌组织与非肿瘤组织中微小RNA种类的差异表达

Differential expression of microRNA species in human gastric cancer versus non-tumorous tissues.

作者信息

Guo Junming, Miao Ying, Xiao Bingxiu, Huan Rong, Jiang Zhen, Meng Dan, Wang Yanjun

机构信息

Ningbo University School of Medicine, Ningbo 315211, Zhejiang Province, China.

出版信息

J Gastroenterol Hepatol. 2009 Apr;24(4):652-7. doi: 10.1111/j.1440-1746.2008.05666.x. Epub 2008 Nov 3.

Abstract

BACKGROUND AND AIM

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.

METHODS

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 microParaflo 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.

RESULTS

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.

CONCLUSION

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.

摘要

背景与目的

微小RNA(miRNA)在肿瘤发生过程中发挥着重要作用。目前尚未见胃癌整体miRNA表达谱的报道。本研究旨在确定胃癌的miRNA表达谱。

方法

首先从原发性胃癌组织及相邻非肿瘤组织中提取总RNA,然后对分离出的小RNA(<300 nt)进行3'端加尾(poly(A))。在μParaflo微流控芯片(美国得克萨斯州休斯顿市LC Sciences公司)上进行杂交。使用特异性标签的Cy3和Cy5染料进行荧光标记杂交检测后,用激光扫描仪采集杂交图像,并用Array-Pro图像分析软件(美国马里兰州银泉市Media Cybernetics公司)进行数字化处理。为验证结果并探究差异表达miRNA的生物学意义,采用免疫组织化学法检测靶基因的差异表达。

结果

非肿瘤组织中表达最高的miRNA为miR-768-3p、miR-139-5p、miR-378、miR-31、miR-195、miR-497和miR-133b。其中,miR-139-5p、miR-497和miR-768-3p首次在非肿瘤组织中被发现。胃癌组织中表达最高的miRNA为miR-20b、miR-20a、miR-17、miR-106a、miR-18a、miR-21、miR-106b、miR-18b、miR-421、miR-340*、miR-19a和miR-658。其中,miR-340*、miR-421和miR-658首次被发现在癌细胞中高表达。发现癌组织中一些靶基因(如Rb和PTEN)的表达降低。

结论

据我们所知,这是关于这些与胃癌相关的miRNA的首次报道。这一新信息可能提示这些miRNA在胃癌诊断中的潜在作用。

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