Department of Medical Sciences, Uppsala University, Uppsala, Sweden.
Mod Pathol. 2013 May;26(5):685-96. doi: 10.1038/modpathol.2012.216. Epub 2013 Jan 18.
Well-differentiated small intestinal neuroendocrine tumors are rare malignancies. They arise from enterochromaffin cells and very little is known about differential microRNA (miRNA) expression. The aim of this study was to identify the miRNA profile of well-differentiated small intestinal neuroendocrine tumors, which may have a critical role in tumor development, progression and potentially develop miRNAs as novel clinical biomarkers. Specimens from two test groups, 24 small intestinal neuroendocrine tumor specimens at different stages of malignancy, are included in this study. Total RNA from the first test group, five primary tumors, five mesentery metastases and five liver metastases was hybridized onto the Affymetrix Genechip miRNA arrays to perform a genome-wide profile. The results were validated by using quantitative real-time PCR (QRT-PCR) and northern blot analyses. We then expanded the investigation to laser capture microdissected small intestinal neuroendocrine tumor cells and immuno-laser capture microdissected normal enterochromaffin cells of the first test group. Furthermore, a second test group, three primary tumors, three mesentery metastases and three liver metastases, was included in the study. Thus, two independent test groups validated the data by QRT-PCR. Moreover, we characterized nine miRNAs, five (miR-96, -182, -183, -196a and -200a), which are upregulated during tumor progression, whereas four (miR-31, -129-5p, -133a and -215) are downregulated. Several online software programs were used to predict potential miRNA target genes to map a number of putative target genes for the aberrantly regulated miRNAs, through an advanced and novel bioinformatics analysis. Our findings provide information about pivotal miRNAs, which may lead to further insights into tumorigenesis, progression mechanisms and novel therapeutic targets recognition.
分化良好的小肠神经内分泌肿瘤是罕见的恶性肿瘤。它们起源于肠嗜铬细胞，关于差异 microRNA（miRNA）表达的了解甚少。本研究的目的是鉴定分化良好的小肠神经内分泌肿瘤的 miRNA 谱，这些 miRNA 可能在肿瘤发生、发展中具有关键作用，并可能将 miRNAs 作为新型临床生物标志物进行开发。本研究纳入了两个实验组的标本，共 24 份处于不同恶性阶段的小肠神经内分泌肿瘤标本。第一个实验组的 5 份原发性肿瘤、5 份肠系膜转移和 5 份肝转移的总 RNA 与 Affymetrix Genechip miRNA 阵列杂交，进行全基因组分析。结果通过定量实时 PCR（QRT-PCR）和 northern blot 分析进行验证。然后，我们将研究扩展到激光捕获微切割的小肠神经内分泌肿瘤细胞和第一个实验组的免疫激光捕获微切割的正常肠嗜铬细胞。此外，第二个实验组纳入了 3 份原发性肿瘤、3 份肠系膜转移和 3 份肝转移的标本。因此，通过 QRT-PCR 验证了两个独立实验组的数据。此外，我们还鉴定了 9 个 miRNA，其中 5 个（miR-96、-182、-183、-196a 和 -200a）在肿瘤进展过程中上调，而 4 个（miR-31、-129-5p、-133a 和 -215）下调。使用了几种在线软件程序来预测潜在的 miRNA 靶基因，以通过先进的新型生物信息学分析来映射异常调节的 miRNA 的一些潜在靶基因。我们的研究结果提供了关键 miRNA 的信息，这些 miRNA 可能进一步深入了解肿瘤发生、进展机制和识别新的治疗靶点。
