摘要/Abstract

摘要： 目的 ·探讨含 V-set域 T细胞激活抑制因子（ V-set domain containing T cell activation inhibitor 1，VTCN1）在结肠癌中对下游长链非编码 RNA（long noncoding RNA，lncRNA）和 mRNA的调控作用。方法 ·利用慢病毒质粒在结肠癌细胞株 SW1116中过表达 VTCN1，抽提 RNA并测序，与阴性对照组比较分析差异表达的 lncRNA和 mRNA，并任意选取差异表达的 lncRNA（3条）和 mRNA（2条）进行实时定量 PCR（qRT-PCR）验证 RNA测序结果的准确性。通过 BLAST2GO和京都基因与基因组百科全书（Kyoto Encyclopedia of Genes and Genomes，KEGG）分析预测这些差异表达 lncRNA和 mRNA的相关功能。利用线上平台 GEPIA18（Gene Expression Profiling Interactive Analysis）分析差异表达 lncRNA与结直肠癌患者生存期的相关性。结果 ·通过 RNA测序在过表达 VTCN1的 SW1116细胞中发现了 167条差异基因，包括 39条 lncRNA和 128条 mRNA。qRT-PCR检验结果与 RNA测序结果变化趋势一致。生物信息学分析结果显示这些受 VTCN1调控的差异基因可能参与了内质网中的蛋白处理、 mRNA监控信号通路。另外，在过表达 VTCN1的结肠癌细胞中明显上调的 3条 lncRNA（DNAJC9-AS1、HCG27和 RP11-339B21.13），同时也是预测结直肠癌患者总生存期的独立因子。结论 ·在结肠癌细胞中 VTCN1对下游多条 lncRNA和 mRNA具有调控作用，这些 lncRNA和 mRNA可能参与了内质网中的蛋白处理和 mRNA监控信号通路。
关键词: 含 V-set域 T细胞激活抑制因子, 结肠癌, 长链非编码 RNA, RNA测序, 生物信息学分析
Abstract:
Objective · To investigate the effect of V-set domain containing T cell activation inhibitor 1 (VTCN1) on long noncoding RNAs (lncRNAs) and mRNAs in colon cancer cells. Methods · VTCN1 was overexpressedlentivirus plasmid in colon cancer cell line SW1116. RNA was extracted and sequenced. The differentially expressed lncRNAs and mRNAs were compared with the negative control group. The accuracy of RNA sequencing was verifiedreal-time quantitative PCR (qRT-PCR) using three differentially expressed lncRNAs and two mRNAs. BLAST2GO and Kyoto Encyclopedia of Genes and Genomes (KEGG) were used to analyze and predict the functions of these differentially expressed lncRNAs and mRNAs. The online platform GEPIA18 (Gene Expression Profiling Interactive Analysis) was used to analyze the correlation between differentially expressed lncRNAs and survival of patients with colorectal cancer. Results · A total of 167 differential genes, i.e., 39 lncRNAs and 128 mRNAs, were identifiedRNA sequencing in VTCN1 overexpressed SW1116 cells. The results of qRT-PCR were consistent with those of RNA sequencing. Bioinformatics analysis showed that these different genes regulatedVTCN1 may be involved in endoplasmic reticulum protein processing and RNA monitoring signaling pathways. In addition, three lncRNAs (DNA JC9-AS1, HCG27, and RP11-339B21.13), which were significantly up-regulated in colorectal cancer cells overexpressing VTCN1, were also independent predictors of overall survival of colorectal cancer patients. Conclusion · In colon cancer cells, VTCN1 regulates several downstream lncRNAs and mRNAs, which may be involved in endoplasmic reticulum protein processing and mRNA monitoring signaling pathways.
Key words: V-set domain containing T cell activation inhibitor 1 (VTCN1), colon cancer, long noncoding RNA (lncRNA), RNA sequencing, bioinformatic analysis


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