Ding Xiangxiang, Zhang Shuai, Li Xiao, Feng Changjiang, Huang Qi, Wang Shaodong, Wang Siwei, Xia Wenjia, Yang Fan, Yin Rong, Xu Lin, Qiu Mantang, Li Ming, Wang Jun
Department of Radiology Jiangsu Cancer Hospital Jiangsu Institute of Cancer Research Nanjing Medical University Affiliated Cancer Hospital Nanjing China.
Department of Thoracic Surgery Jiangsu Key Laboratory of Molecular and Translational Cancer Research Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research Nanjing Medical University Affiliated Cancer Hospital Nanjing China.
FEBS Open Bio. 2018 Feb 21;8(4):544-555. doi: 10.1002/2211-5463.12397. eCollection 2018 Apr.
Noncoding RNA play important roles in various biological processes and diseases, including cancer. The expression profile of circular RNA (circRNA) has not been systematically investigated in lung adenocarcinoma (LUAD). In this study, we performed genomewide transcriptome profiling of coding genes, long noncoding RNA (lncRNA), and circRNA in paired LUAD and nontumor tissues by ribosomal RNA-depleted RNA sequencing. The detected reads were first mapped to the human genome to analyze expression of coding genes and lncRNA, while the unmapped reads were subjected to a circRNA prediction algorithm to identify circRNA candidates. We identified 1282 differentially expressed coding genes in LUAD. Expression of 19 023 lncRNA was detected, of which 244 lncRNAs were differentially expressed in LUAD. AFAP1-AS1, BLACAT1, LOC101928245, and FENDRR were most differentially expressed lncRNAs in LUAD. Also identified were 9340 circRNA candidates with ≥ 2 backspliced, including 3590 novel circRNA transcripts. The median length of circRNA was ~ 530 nt. CircRNA are often of low abundance, and more than half of circRNAs we identified had < 10 reads. Agarose electrophoresis and Sanger sequencing were used to confirm that four candidate circRNA were truly circular. Our results characterized the expression profile of coding genes, lncRNA, and circRNA in LUAD; 9340 circRNAs were detected, demonstrating that circRNA are widely expressed in LUAD.
The raw RNA sequencing data have been submitted to Gene Expression Omnibus (GEO) database and can be accessed with the ID GEO: http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE104854.
非编码RNA在包括癌症在内的各种生物过程和疾病中发挥重要作用。环状RNA(circRNA)的表达谱尚未在肺腺癌(LUAD)中进行系统研究。在本研究中,我们通过核糖体RNA去除RNA测序对配对的LUAD和非肿瘤组织中的编码基因、长链非编码RNA(lncRNA)和circRNA进行了全基因组转录组分析。检测到的 reads 首先映射到人类基因组以分析编码基因和lncRNA的表达,而未映射的 reads 则经过circRNA预测算法以识别circRNA候选物。我们在LUAD中鉴定出1282个差异表达的编码基因。检测到19023个lncRNA的表达,其中244个lncRNA在LUAD中差异表达。AFAP1-AS1、BLACAT1、LOC101928245和FENDRR是LUAD中差异表达最明显的lncRNA。还鉴定出9340个候选circRNA,其反向剪接数≥2,包括3590个新的circRNA转录本。circRNA的中位数长度约为530 nt。circRNA通常丰度较低,我们鉴定出的circRNA中超过一半的 reads < 10。使用琼脂糖凝胶电泳和桑格测序来确认四个候选circRNA是真正的环状。我们的结果描绘了LUAD中编码基因、lncRNA和circRNA的表达谱;检测到9340个circRNA,表明circRNA在LUAD中广泛表达。
原始RNA测序数据已提交至基因表达综合数据库(GEO),可通过ID GEO:http://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE104854访问。