| 摘要: |
| 为鉴定MD淋巴瘤转化细胞中gga-miR-130b-3p 调控的肿瘤相关基因，本研究在MDV转化的鸡淋巴细胞系MDCC-MSB1中过表达该miRNA，转染48 h后收集并提取RNA，利用高通量测序获得过表达gga-miR-130b-3p组和阴性对照组（Negative control， NC）的转录组数据，筛选这些数据中的差异表达基因（Differential expressed genes， DEGs），对DEGs进行基因本体 GO功能富集分析、信号通路分析以及基因集富集分析，筛选过表达gga-miR-130b-3p前后基因表达变化可能参与的信号通路，并对两组中的差异可变剪切进行分析。结果表明：1）与NC组相比，过表达gga-miR-130b-3p组中共鉴定出117个DEGs，其中83个DEGs显著上调，34个DEGs显著下调。其中MCM10、KCNA3、PTK2、FGL2、GPAM、BMP4、LOXL3、DDO和KRAS等基因可能影响MD肿瘤转化。2）DEGs注释到46个GO条目中，其中生物过程包含免疫系统过程等在内的22个条目，分子功能包括10个条目，细胞组分包括14个条目。3）mimics NC和mimics转染组中的DEGs以及微效基因富集到7个与肿瘤发生相关的通路，其中主要是通过甲状腺激素信号途径、胆碱代谢等通路发挥抑癌作用。4）过表达gga-miR-130b-3p后差异可变剪切类型最多的是外显子跳跃（Exon skipped，ES），其次是内含子滞留（Intron retained，IR），最少的是外显子互斥（Mutually exclusive exon，MXE）。综上， gga-miR-130b-3p高表达会引起MSB1细胞中的部分基因差异表达，这些DEGs通过抑癌信号通路抑制MD的肿瘤发展进程，该研究可为解析miRNA在MD肿瘤转化过程中的作用机制提供理论依据。 |
| 关键词: gga-miR-130b-3p MD肿瘤相关基因 转录组测序 生物信号通路 |
| DOI：10.11841/j.issn.1007-4333.2024.07.04 |
| 投稿时间：2024-03-03 |
| 基金项目:国家自然科学基金项目 （32002160，32172816）；安徽省教育厅青年拔尖人才项目；安徽省高校科研项目计划自然科学重点项目 （2023AH051872）；安徽省重点研究与开发计划项目 （202204c06020074）； 安徽科技学院人才引进项目 （DKYJ201901）；安徽省研究生创新创业实践项目（2023cxcysj175）；大学生创新创业训练计划项目 （202110879059， S202210879200，S202210879194，X202310879019） |
|
| Effects of gga-miR-130b-3p on the expression of tumour-related genes in chicken MDCC-MSB1 cells |
|
LI Jiale, JIN Tao, ZHU Guozhi, CHEN Shenjun, YANG Lin, LIU Xinyu, LI Shenghe, ZHAO Chunfang*
|
| (Anhui Provincial Key Laboratory of Animal Nutrition Regulation and Health /College of Animal Science， Anhui Science and Technology University， Chuzhou 233100，China) |
| Abstract: |
| To identify tumour-associated genes regulated by gga-miR-130b-3p in MD lymphoma-transformed cells， in the study， the miRNA mimics was overexpressed in MDV-transformed chicken lymphoblastoid cell line MDCC-MSB1， and cells were collected and RNA extracted 48 h after transfection. The transcriptome data were obtained for the overexpression of gga-miR-130b-3p group and the negative control （NC） group using high-throughput sequencing. Screening these data for differentially expressed genes （DEGs）， functional enrichment analysis of DEGs for Gene ontology （GO）， signaling pathway analysis and Gene Set Enrichment Analysis （GSEA） were performed to screen the signaling pathways that may be involved in the changes in gene expression before and after overexpression of gga-miR-130b-3p and to analyze the differentially alternative splicing in the two groups. The results showed that： 1） Compared to the NC group， a total of 117 DEGs were identified in the overexpression gga-miR-130b-3p group， of which 83 DEGs were significantly up-regulated and 34 DEGs were significantly down-regulated. Among them， MCM10， KCNA3， PTK2， FGL2， GPAM， BMP4， LOXL3， DDO and KRAS may affect MD tumor transformation. 2） DEGs were annotated into 46 GO entries， of which 22 entries including immune system processes for biological processes， 10 entries for molecular functions， and 14 entries for cellular components. 3） DEGs in the mimics NC and mimics transfected groups as well as genes of small effect were enriched to seven pathways associated with tumourigenesis， which mainly exerted oncogenic effects through the thyroid hormone signaling pathway， choline metabolism and other pathways. 4） The most differentially alternative splicing type after overexpression of gga-miR-130b-3p was exon skipped （ES）， followed by intron retained （IR）， and the least was mutually exclusive exon （MXE）. Taken together， high expression of gga-miR-130b-3p can cause differential expression of some genes in MSB1 cells， and these DEGs inhibit tumour progression in MD through oncogenic signaling pathways. This study may provide a theoretical basis for analyzing the mechanism of gga-miR-130b-3p in MD tumour transformation process. |
| Key words: gga-miR-130b-3p Marek's disease transcriptome sequencing biosignaling pathway |
