一种与肝细胞癌相关的新型长链非编码RNA基因的克隆和功能表征

Pub Date : 2014-01-01 DOI:10.3724/SP.J.1206.2012.00613
Gong Zj, Ming Zhou, Peng Sp, J. Ma, Xiaoyan Li, Hongbin Huang, Hongbin Huang, Y. Li, Xiaoling Li, H. Bo, Zeng Zy, Strong, Gui-yuan Li, Xiang Jj, W. Xiong, F. Wei, Zhang Wl, Liao Qj, K. Tang, Song Yl
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引用次数: 14

摘要

最近,我们利用基于下一代测序(NGS)技术的rna -测序(RNA-Seq)策略对肝细胞癌活检组织和正常肝组织的转录组进行了测序,并在肝细胞癌活检组织的11q13.1染色体上发现了几个相邻的高RNA-Seq信号峰,而在正常对照组织中则没有。在这个染色体区域,没有鉴定到特征基因,这意味着这些RNA-Seq峰可能代表一个或多个新基因。进一步的研究证实,这些RNA-Seq峰是由一个新基因转录的。通过克隆该新基因的全长,我们发现该新基因转录了多个剪接异构体,最长的异构体为3 562 bp。然后将12个具有代表性的RNA异构体存入国家生物技术信息中心(NCBI)的GenBank数据库,并为这些异构体创建了从KC136297到KC136308的GenBank id。在该基因的所有转录本中均未发现显著的开放阅读片段(ORF),这表明该基因可能编码长链非编码rna (lncRNAs)。为了进一步阐明该lncRNA基因潜在的转录调控机制,我们利用生物信息学工具对lncRNA基因上游序列的启动子进行了预测,发现在lncRNA基因转录起始位点719 ~ 469 bp处存在1个潜在启动子,并且在启动子区域存在7个Sp1、1个STAT5和1个EGR1转录因子结合位点。lncRNA基因在肝癌发生发展中的分子机制值得进一步研究。
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Cloning and Functional Characterization of a Novel Long Non-coding RNA Gene Associated With Hepatocellular Carcinoma
Recently, we sequenced the transcriptomes of a hepatocellular carcinoma biopsy and a normal liver tissue using the RNA-Sequencing(RNA-Seq) strategy based on the Next Generation Sequencing(NGS) technique, and identified several adjacent high RNA-Seq signal peaks on chromosome 11q13.1 in the hepatocellular carcinoma biopsy, while not in the normal control tissue. In this chromosome region, there is no characterized genes have been identified, implying that these RNA-Seq peaks may represent one or more novel genes. Further study was confirmed that these RNA-Seq peaks were transcribed by one novel gene. Through cloning the full length of this novel gene, we found that this novel gene transcribed many splicing isoforms, and the longest isoform is 3 562 bp. Then we deposited twelve representative RNA isoforms into the GenBank database of the National Center for Biotechnology Information(NCBI), and created the GenBank IDs from KC136297 to KC136308 for these isoforms. None significant open reading fragment(ORF) was found in any transcripts of this novel gene, implying that this gene may encodes long non-coding RNAs(lncRNAs). To further elucidate the potential transcriptional regulation mechanism of this lncRNA gene, we predicted the promoter from the upstream sequence of the lncRNA gene using bioinformatic tools, and found that there is one potential promoter in-719 to-469 bp from the transcript start site of the lncRNA gene, and there are seven Sp1, one STAT5 and one EGR1 transcription factor binding sites in the promoter region. The molecular mechanisms of the lncRNA gene in carcinogenesis and progression of hepatocellular carcinoma are worthful for further investigation.
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