Chen Dong, Sun Qiang, Cheng Xiaofei, Zhang Lufei, Song Wei, Zhou Dongkai, Lin Jianjiang, Wang Weilin
Department of Colorectal Surgery, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.
Division of Hepatobiliary and Pancreatic Surgery, Department of Surgery, First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, China.
Cancer Med. 2016 Jul;5(7):1629-39. doi: 10.1002/cam4.738. Epub 2016 May 11.
The liver is the most frequent site of metastasis in colorectal cancer (CRC), in which long noncoding RNAs (lncRNAs) may play a crucial role. In this study, we performed a genome-wide analysis of lncRNA expression to identify novel targets for the further study of liver metastasis in CRC. Samples obtained from CRC patients were analyzed using Arraystar human 8 × 60K lncRNA/mRNA v3.0 microarrays chips to find differentially expressed lncRNAs and mRNAs. The results were confirmed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The differentially expressed lncRNAs and mRNAs were identified through fold change filtering. Gene ontology (GO) and pathway analyses were performed using standard enrichment computational methods. In the CRC tissues from patients with liver metastasis, 2636 lncRNAs were differentially expressed, including 1600 up-regulated and 1036 down-regulated over two-fold compared with the CRC tissues without metastasis. Among the 1584 differentially expressed mRNAs, 548 were up-regulated and 1036 down-regulated. GO and pathway analysis of the up-regulated and down-regulated mRNAs yielded different results. The up-regulated mRNAs were associated with single-organism process (biological process), membrane part (cellular component), and transporter activity (molecular function), whereas the down-regulated mRNAs were associated with cellular process, membrane, and binding, respectively. In the pathway analysis, 27 gene pathways associated with the up-regulated mRNAs and 51 gene pathways associated with the down-regulated mRNAs were targeted. The significant changes in NQO2 (NM_000904) mRNA and six associated lncRNAs were selected for validation by qRT-PCR. Aberrantly expressed lncRNAs may play an important role in the liver metastasis of CRC. The further study can provide useful insights into the biology and, ultimately, the prevention of liver metastasis.
肝脏是结直肠癌（CRC）最常见的转移部位，长链非编码RNA（lncRNAs）可能在其中发挥关键作用。在本研究中，我们对lncRNA表达进行了全基因组分析，以确定CRC肝转移进一步研究的新靶点。使用Arraystar人类8×60K lncRNA/mRNA v3.0微阵列芯片分析从CRC患者获得的样本，以发现差异表达的lncRNAs和mRNAs。结果通过定量逆转录-聚合酶链反应（qRT-PCR）得到证实。通过倍数变化筛选确定差异表达的lncRNAs和mRNAs。使用标准富集计算方法进行基因本体（GO）和通路分析。在有肝转移患者的CRC组织中，与无转移的CRC组织相比，有2636个lncRNAs差异表达，其中1600个上调，1036个下调超过两倍。在1584个差异表达的mRNAs中，548个上调，1036个下调。上调和下调mRNAs的GO和通路分析产生了不同的结果。上调的mRNAs分别与单细胞过程（生物学过程）、膜部分（细胞成分）和转运蛋白活性（分子功能）相关，而下调的mRNAs分别与细胞过程、膜和结合相关。在通路分析中，分别靶向了与上调mRNAs相关的27个基因通路和与下调mRNAs相关的51个基因通路。选择NQO2（NM_000904）mRNA和6个相关lncRNAs的显著变化通过qRT-PCR进行验证。异常表达的lncRNAs可能在CRC肝转移中起重要作用。进一步的研究可为CRC肝转移的生物学机制乃至预防提供有用的见解。
