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整合 RNA-Seq 数据的分析和 qRT-PCR 验证鉴定结直肠癌中常见失调的基因。

Identification of commonly dysregulated genes in colorectal cancer by integrating analysis of RNA-Seq data and qRT-PCR validation.

机构信息

Department of Oncology, The First affiliated Hospital of PLA General Hospital, Beijing, China.

Beijing Yangshen Bioinformatic Technology, Beijing, China.

出版信息

Cancer Gene Ther. 2015 May;22(5):278-84. doi: 10.1038/cgt.2015.20. Epub 2015 Apr 24.

Abstract

The progression of colorectal cancer (CRC) is a multistep process and metastatic CRC is always incurable; consequently, CRC is the leading cause of cancer-related deaths. There is therefore an urgent need for identifying useful biomarkers with enough sensitivity and specificity to detect this disease at early stages, which will significantly reduce the mortality for this malignancy. In this study, we performed an integrating analysis of different RNA-Seq data sets to find new candidate biomarkers for diagnosis, prognosis and as therapeutic targets for this malignancy, as well as to elucidate the molecular mechanisms of CRC carcinogenesis. We identified 883 differentially expressed genes (DEGs) across the studies between CRC and normal control (NC) tissues by combining five RNA-Seq data sets. Gene function analysis revealed high correlation with carcinogenesis. The top 10 most significantly DEGs were further evaluated by quantitative real-time polymerase chain reaction (qRT-PCR) in both rectal cancer (RC) and colon cancer (CC), and the results matched well with integrating data, suggesting that the method of integrating analysis of different RNA-seq data sets is acceptable. Therefore, integrating analysis of different RNA-seq data sets may be a useful way to overcome the limitation of small sample size in a single RNA-seq study. In addition, our study showed that some genes, such as SIM2, ADAMTS6, FOXD4L4 and DNAH5, may have an important role in the development of CRC, which could be applied for diagnosis, prognosis and as therapy for this malignancy. Our findings would also help to understand the pathology of CRC.

摘要

结直肠癌(CRC)的进展是一个多步骤的过程,转移性 CRC 总是无法治愈;因此,CRC 是癌症相关死亡的主要原因。因此,迫切需要确定具有足够灵敏度和特异性的有用生物标志物,以便在早期检测到这种疾病,这将显著降低这种恶性肿瘤的死亡率。在这项研究中,我们对不同的 RNA-Seq 数据集进行了综合分析,以发现新的候选生物标志物,用于诊断、预后和作为这种恶性肿瘤的治疗靶点,并阐明 CRC 癌变的分子机制。我们通过结合五个 RNA-Seq 数据集,在 CRC 和正常对照(NC)组织之间的不同研究中发现了 883 个差异表达基因(DEGs)。基因功能分析显示与癌变高度相关。前 10 个差异最显著的 DEGs 进一步通过定量实时聚合酶链反应(qRT-PCR)在直肠癌(RC)和结肠癌(CC)中进行评估,结果与综合数据吻合较好,表明整合不同 RNA-seq 数据集的分析方法是可以接受的。因此,整合不同 RNA-seq 数据集的分析可能是克服单个 RNA-seq 研究中样本量小的局限性的一种有用方法。此外,我们的研究表明,一些基因,如 SIM2、ADAMTS6、FOXD4L4 和 DNAH5,可能在 CRC 的发展中具有重要作用,可用于诊断、预后和治疗这种恶性肿瘤。我们的发现还有助于理解 CRC 的病理学。

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