实用医学杂志 ›› 2023, Vol. 39 ›› Issue (9): 1079-1085.doi: 10.3969/j.issn.1006⁃5725.2023.09.003

• 基础研究 • 上一篇    下一篇

基于转录组测序技术筛选NSUN2在卵巢癌细胞的基因差异表达

何婉珊1,2 洪小山2 陈冠桥1 陈斌1 文斌2 林育3 韦之富4 罗喜平1,2    

  1. 1 广州医科大学(广州 511436);2 广东省妇幼保健院妇科(广州 511400);3 南方医科大学(广州 510515); 4 暨南大学附属顺德医院妇科(广东佛山 528000)

  • 出版日期:2023-05-10 发布日期:2023-05-10
  • 通讯作者: 罗喜平 E⁃mail:luoxiping333@126.com
  • 基金资助:
    广州市科技计划项目(编号:202002030174)

Screening of gene differential expression of NSUN2 in ovarian cancer cells based on transcriptome sequenc⁃ ing technology

HE Wanshan* ,HONG Xiaoshan,CHEN Guanqiao,CHEN Bin,WEN Bin,LIN Yu,WEI Zhifu, LUO Xiping   

  1. Guangzhou Medical University,Guangzhou 511436,China;*Department of Gynecology, the Guang⁃ dong Women and Children Hospital, Guangzhou 511400, China

  • Online:2023-05-10 Published:2023-05-10
  • Contact: LUO Xiping E⁃mail:luoxiping333@126.com

摘要:

目的 应用 RNA⁃seq 技术研究 NSUN2 对卵巢癌细胞株 A2780 基因表达的影响,为阐明 NSUN2 在卵巢癌症发生发展中的作用提供理论依据。方法 慢病毒介导的 RNA 干扰技术抑制 A2780 细 胞中 NSUN2 的表达。以转染空载体的细胞为对照,使用 RNA⁃seq 技术对干扰细胞株进行基因谱高通量测 序分析,筛选出差异表达的基因。并通过 KEGG Pathway、GO 分析和 GSEA 富集分析,探索 NSUN2 调控的 差异基因主要参与的相关信号通路和生物学过程。结果 与对照组细胞相比,NSUN2 敲低的 A2780 细胞 株有 1 642 个差异表达的基因,其中上调基因有 1 020 个,下调基因有 622 个;通过 GO 分析、KEGG Pathway 和 GSEA 富集分析发现差异表达基因主要富集在 hedgehog 信号通路、P53 等信号通路上,参与了病毒转录、 翻译起始和调节等生物过程;通过对 P53 信号通路的相关基因进一步分析,筛选出与改通路相关的 3 个关 键基因(ZMAT3、EI24 和 CCND2),RT⁃qPCR 验证其表达结果与测序结果相符。结论 A2780 sh⁃NSUN2 组 与 A2780 sh⁃NC 组的基因表达谱间存在明显的差异,其差异基因主要富集于 P53 信号通路。在卵巢癌中 NSUN2可能通过调控P53信号通路相关因子的表达,从而调控了卵巢癌的发生发展。

关键词: NSUN2, 卵巢癌, 转录组测序技术, P53信号通路 ,

Abstract:

Objective To investigate the effect of gene expression by NSUN2 in ovarian cancer cell line A2780 by RNA ⁃seq technology,and to provide a theoretical basis for clarifying the role of NSUN2 in the occur⁃ rence and progression of ovarian cancer. Methods Expression of NSUN2 in A2780 cells was knocked down by lentivirus⁃mediated RNA interference technique. The cells transfected with empty vector were used as control,and the interfering cell lines were screened for differentially expressed genes by high⁃throughput sequencing using RNA⁃ seq technology. KEGG Pathway,GO analysis and GSEA enrichment analysis were used to explore the related sig⁃ naling pathways and biological processes of NSUN2⁃regulated differential genes. Results Compared with the con⁃ trol cells,there were 1 642 differentially expressed genes in the A2780 cell line with NSUN2 knockdown,includ⁃ ing 1 020 up⁃regulated genes and 622 down⁃regulated genes. Through GO analysis,KEGG Pathway and GSEA en⁃ richment analysis,it was found that differentially expressed genes were mainly enriched in hedgehog signaling path⁃ way and P53 signaling pathway,and were involved in viral transcription,translation initiation and regulation. Three key genes related to the P53 signaling pathway(ZMAT3,EI24 and CCND2)were screened out through fur⁃ ther analysis of the genes related to the p53 signaling pathway. The expression results were verified by RT ⁃qPCR and the sequencing results were consistent. Conclusion There are significant differences in gene expression pro⁃ files between A2780 sh⁃NSUN2 group and A2780 sh⁃NC group,and the differential genes are mainly concentrated in P53 signaling pathway. In ovarian cancer,NSUN2 may regulate the occurrence and development of ovarian can⁃ cer by regulating the expression of P53 signaling pathway related factors.

Key words:

NSUN2, ovarian cancer, RNA?seq technology, P53 signaling pathway