基于RNA-Seq技术筛选的功能性融合转录本TSNAX-DISC1与子宫内膜癌发生发展的关联研究

发布时间：2018-09-01 19:39

【摘要】：作为一种特殊的个体突变，融合基因是由两个相邻基因的全部或一部分序列相互融合，通过基因间剪接从而形成的一种新的转录物。越来越多的研究表明，融合基因与多种肿瘤的形成和发展密切相关，，但是与子宫内膜癌的相关性研究还尚未报道。本研究采用Ilumina HiSequence2000测序平台对9对子宫内膜癌和癌旁组织进行100bp双末端转录本测序检测差异表达的融合基因。结果发现位于子宫内膜癌易感区域1q42.2内的融合转录本TSNAX-DISC1在子宫内膜癌和癌旁组织中存在显著的表达差异，该融合转录本在多数癌组织中的表达水平显著上调。利用逆转录聚合酶链反应（reverse transcription PCR，RT-PCR）的方法，我们发现融合转录本TSNAX-DISC1并非通过染色体重排的方式形成，而是由相邻基因TSNAX和DISC1的基因间剪接导致的。进一步的数据分析显示，CCCTC-结合因子（CTCF）在TSNAX和DISC1基因间的绝缘子元件上的结合强度与该融合转录本的表达水平呈负相关。位于TSNAX和DISC1基因间的长链非编码RNAlincRNA-NR_034037，通过干扰绝缘子与CTCF的相互作用调控融合转录本TSNAX-DISC1的形成以及TSNAX/DISC1的表达水平。另外，荧光素酶报告基因以及染色质免疫共沉淀（ChIP）实验结果显示，异常表达的TSNAX可显著抑制TRAX、Translin和SF-1三者形成的蛋白复合体在类固醇激素合成急性调控蛋白（StAR）基因启动子上的募集，调节StAR基因的转录活性。这些研究结果表明，lincRNA-NR_034037可能通过调控融合转录本TSNAX-DISC1的形成，参与子宫内膜癌的病理过程。
[Abstract]:As a special individual mutation, fusion gene is a new transcription material, which is formed by the fusion of all or part of the sequence of two adjacent genes by splicing between genes. More and more studies have shown that the fusion gene is closely related to the formation and development of multiple tumors, but the correlation with endometrial carcinoma has not been reported. In this study, Ilumina HiSequence2000 sequencing platform was used to detect the differentially expressed fusion genes in 9 pairs of endometrial carcinoma and adjacent tissues by 100bp double terminal transcripts sequencing. The results showed that there was a significant difference in the expression of fusion transcript TSNAX-DISC1 in endometrial carcinoma and paracancerous tissues, and the expression level of the fusion transcript was up-regulated in most cancer tissues. By reverse transcriptase polymerase chain reaction (reverse transcription PCR,RT-PCR), we found that the fusion transcripts TSNAX-DISC1 were not formed by chromosome rearrangement, but by splicing between adjacent genes TSNAX and DISC1. Further data analysis showed that the binding intensity of CCCTC- binding factor (CTCF) on insulator elements between TSNAX and DISC1 genes was negatively correlated with the expression level of the fusion transcript. The long strand non-coding RNAlincRNA-NR_034037, located between TSNAX and DISC1 genes regulates the formation of TSNAX-DISC1 and the expression of TSNAX/DISC1 by interfering with the interaction between insulators and CTCF. In addition, luciferase reporter gene and chromatin immunoprecipitation (ChIP) results showed that, The abnormal expression of TSNAX could significantly inhibit the recruitment of protein complexes formed by TRAX,Translin and SF-1 on the promoter of steroid hormone synthesis acute regulatory protein (StAR) gene and regulate the transcriptional activity of StAR gene. These results suggest that lincRNA-NR034037 may be involved in the pathological process of endometrial carcinoma by regulating the formation of fusion transcripts TSNAX-DISC1.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

【共引文献】