| 朱豆豆,陈 利,徐 园,李 凯,周宇荀,肖君华.Srsf1敲低对GT1-7细胞基因表达谱的影响[J].,2019,19(14):2614-2619 |
| Srsf1敲低对GT1-7细胞基因表达谱的影响 |
| Effect on Gene Expression Profile of Srsf1 Knockdown in GT1-7 Cells |
| 投稿时间：2018-12-07 修订日期：2018-12-30 |
| DOI：10.13241/j.cnki.pmb.2019.14.003 |
| 中文关键词: 精氨酸/丝氨酸富集的剪接因子1 GT1-7细胞 转录组测序 |
| 英文关键词: Srsf1 GT1-7 RNA-Seq |
| 基金项目:国家自然科学基金项目（31772550）；上海市科委基金资助项目（14140900502） |
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| 中文摘要: |
| 摘要 目的：本研究通过RNA-seq技术分析Srsf1基因表达敲低的GT1-7细胞（knock down , KD）和野生型GT1-7细胞（wide type, WT）的表达谱和可变剪接事件，研究Srsf1敲低对GT1-7细胞基因表达谱的影响。方法：利用实验室现有的Srsf1基因表达敲低的GT1-7细胞（SRSF1-KD）和野生型GT1-7细胞分别抽提RNA，做RNA-seq数据分析，利用rMATS软件分析两株细胞内可变剪接事件的变化，确定Srsf1调控的下游关键基因。结果：结果显示共有875个基因表达存在显著性差异，对这些基因进行KEGG通路分析，发现?酌-氨基丁酸能突触、PI3K-Akt信号通路、MAPK信号通路等与性发育相关的通路均受到影响。此外，P53通路也受到Srsf1敲低的影响。利用rMATS软件进行可变剪接事件分析，显示共发生839个可变剪接事件，涉及719个基因，进行KEGG通路分析发现与性发育相关的MAPK信号通路受到影响。结论：成功检测了GT1-7细胞和Srsf1基因敲低的GT1-7细胞表达谱，通过分析基因表达差异以及可变剪接事件，证明了Srsf1对于PI3K-Akt信号通路、MAPK信号通路、γ-氨基丁酸能突触信号通路、p53通路的调控作用，并且这些信号通路提示我们Srsf1可能更多的通过非可变剪接方式影响性发育相关基因的表达，而非可变剪接方式，从而为更深入的性发育研究提供了理论基础。 |
| 英文摘要: |
| ABSTRACT Objective: In this paper, we analyzed the expression profiles and alternative splicing events of GT1-7 cells with knockdown of Srsf1 (knock down, KD) and wild type GT1-7 cells (wide type, WT) by RNA-seq technology, to research the effect of the Srsf1 knockdown on gene expression profiles in GT1-7 cells. Methods: RNA was extracted from GT1-7 cells with knockdown of Srsf1 (knock down, KD) and wild type GT1-7 cells (wide type, WT) saved in the laboratory for RNA-seq program. Differentially expressed genes were analyzde after processing RNA-seq raw data by bioinformatics methods. Changes in alternative splicing events of two cells were analyzed by rMATS software to identify downstream key genes regulated by Srsf1. Results: The results showed that there were significant differences in the expression level of 875 genes. The KEGG pathway analysis of these differentially expressed genes revealed that several known sexual development related pathways including γ-aminobutyric acid synaptic signaling pathway, PI3K-Akt signaling pathway, MAPK signaling pathway were all affected. In addition, the P53 signaling pathway was also affected by the knockdown of Srsf1. Analysis of alternative splicing events using rMATS software revealed 839 alternative splicing events involving 719 genes. The KEGG pathway analysis of these genes revealed that MAPK signaling pathways associated with sexual development were affected. Conclusion: The expression profiles of GT1-7 cells and Srsf1 knockdown GT1-7 cells were successfully detected by means of RNA-seq techonology. By using bioinformatics methods to analyze gene expression differences and alternative splicing events, it was demonstrated that Srsf1 is involved in PI3K-Akt signaling pathway, MAPK signaling pathway and γ-amino Butyric acid signaling pathway and p53 signaling pathway, and these signaling pathways suggest that Srsf1 may affect the expression of sexual development-related genes more by non-alternative splicing, rather than alternative splicing, thus our results provide a theoretical basis for deepening study on sexual development. |
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