DNA损伤诱导的tsRNA在HepG2细胞中的鉴定和功能研究  

Identification and functional study of DNA damage-induced tsRNAs in HepG2 cells

作　　者：陈观子 洪甫阳 姜袁昊 张裕珍 揭育胜[1] CHEN Guanzi;HONG Fuyang;JIANG Yuanhao;ZHANG Yuzhen;JIE Yusheng(Department of Infectious Diseases,Third Affiliated Hospital of Sun Yat-sen University,Guangzhou 510630,China)

机构地区：[1]中山大学附属第三医院感染科,广东广州510630

出　　处：《中国病理生理杂志》2025年第4期688-695,共8页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金面上项目(No.2021A1515012617)。

摘　　要：目的:鉴定人肝母细胞瘤HepG2细胞中响应DNA损伤的转运RNA衍生的小RNA(transfer RNAderived small RNA,tsRNA)的表达特征,并研究其潜在功能。方法:本研究基于配对的HepG2细胞和敲除TP53基因的HepG2细胞,采用阿霉素(adriamycin,ADR)成功构建DNA损伤的细胞模型,并进行小非编码RNA的转录组分析,系统地鉴定一批响应ADR并参与p53调节的tsRNA,利用京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes,KEGG)进行了功能富集分析。此外,经沉默目标tsRNA基因表达后,通过CCK-8实验和平板集落形成实验初步证实了目标tsRNA在HepG2细胞模型中的生物学功能。结果:DNA损伤可诱导一批参与p53调节的tsRNA,其中tRF-5-1(tRF-5_tRNA-Gly-TCC-2-1)和tRF-i-1(tRF i_tRNA-Tyr-GTA-11-1)在HepG2细胞中的表达上调最为显著(P<0.05)。沉默tRF-5-1或tRF-i-1基因可抑制HepG2细胞的增殖活力(P<0.05)。结论:HepG2细胞模型中可以鉴定一组响应DNA损伤的tsRNA,且tsRNA可以促进HepG2细胞的增殖活力,提示tsRNA在肝脏细胞的恶性增殖中可能扮演重要角色。AIM:To identify the expression characteristics of transfer RNA-derived small RNAs(tsRNAs)responding to DNA damage in human hepatoblastoma HepG2 cells,and to investigate their potential functions.METHODS:Based on paired HepG2 cells and TP53 gene knockout HepG2 cells,we successfully constructed a DNA damage cellular model using adriamycin(ADR).Transcriptome analysis of small noncoding RNAs was performed to systematically identify a set of tsRNAs responding to ADR and involved in p53 regulation.Functional enrichment analysis was conducted using the Kyoto Encyclopedia of Genes and Genomes(KEGG).Additionally,after silencing the expression of target tsRNA genes,the biological functions of these tsRNAs in HepG2 cells were initially confirmed through CCK-8 assay and plate colony formation assay.RESULTS:DNA damage induced a set of tsRNAs involved in p53 regulation,among which tRF-5-1(tRF-5_tRNA-Gly-TCC-2-1)and tRF-i-1(tRF i_tRNA-Tyr-GTA-11-1)showed the most significant up-regulation in HepG2 cells(P<0.05).Silencing of either tRF-5-1 or tRF-i-1 gene inhibited the proliferative activity of HepG2 cells(P<0.05).CONCLUSION:A group of tsRNAs responding to DNA damage can be identified in HepG2 cell model,and tsRNAs can promote the proliferative activity of HepG2 cells,suggesting that tsRNAs may play an important role in the development and progression of liver malignancies.

关 键 词：DNA损伤 转运RNA衍生的小RNA P53蛋白 HEPG2细胞 

分 类 号：R575[医药卫生—消化系统] R735.7[医药卫生—内科学] R363[医药卫生—临床医学]