鉴定顺铂对人肝癌细胞系转录物组的影响

doi:10.16352/j.issn.1001-6325.2024.03.0352

基础医学与临床 ›› 2024, Vol. 44 ›› Issue (3): 352-360.doi: 10.16352/j.issn.1001-6325.2024.03.0352

鉴定顺铂对人肝癌细胞系转录物组的影响

郭鑫, 冀梦蝶, 王琦, 李雪媛, 陈阳^*

中国医学科学院北京协和医学院基础医学研究所生物化学与分子生物学系重大疾病共性机制研究全国重点实验室,北京 100005

收稿日期:2023-11-07 修回日期:2023-12-27 出版日期:2024-03-05 发布日期:2024-02-22
通讯作者: ^*:yc@ibms.pumc.edu.cn
基金资助:
中国医学科学院中央级公益性科研院所基本科研业务费(2021-RC310-007)

Identification of the effect of cisplatin on the transcriptome of human hepatocellular carcinoma cell lines

GUO Xin, JI Mengdie, WANG Qi, LI Xueyuan, CHEN Yang^*

State Key Laboratory of Common Mechanism Research for Major Diseases, Department of Biochemistry and Molecular Biology, Institute of Basic Medical Sciences, CAMS & PUMC, Beijing 100005, China

Received:2023-11-07 Revised:2023-12-27 Online:2024-03-05 Published:2024-02-22
Contact: ^*:yc@ibms.pumc.edu.cn

摘要/Abstract

摘要： 目的通过不同浓度顺铂(CDDP)处理人肝癌细胞系并进行转录物组测序分析,旨在揭示顺铂处理对肝癌细胞转录水平的影响。方法使用终浓度为 0、20、50、100和200 μmol/L的顺铂处理肝癌细胞系HepG2、Huh7 12 h后进行细胞活性检测、免疫荧光和转录物组测序(RNA-seq),并进行差异表达(DEG)、KEGG及蛋白质相互作用网络分析。结果顺铂处理后HepG2、Huh7细胞活性下降,且DNA损伤增多,不同顺铂浓度处理下两种细胞系中共同上调的基因共有59个,共同下调的基因有81个。共同上调的基因主要富集在肿瘤发生发展相关通路,共同下调的81个基因主要富集在Rap1信号通路、Ras信号通路、调控干细胞多能性的信号通路、轴突的指导和黏附连接等相关通路。分析共同上下调基因的蛋白质相互作用网络中关键节点的生存预后,Jun原癌基因,AP-1转录因子(JUN)的高表达与患者生存期的延长呈显著相关,生长停滞DNA损伤可诱导蛋白α(GADD45A)的低表达与患者生存期的延长呈显著相关。结论揭示了肝癌细胞在顺铂处理下的共性转录物变化。JUN和GADD45A的表达差异可能是耐药机制的关键节点,也为临床治疗提供了重要的预后指标。

关键词: 肝癌, 转录物组测序, 顺铂, 基因转录

Abstract: Objective To investigate the effect of cisplatin treatment on the transcriptional level of human liver cancer cells by conducting transcriptome sequencing analysis after treating human liver cancer cell lines with different concentrations of cisplatin (CDDP). Methods Liver cancer cell lines HepG2 and Huh7 were incubated with cisplatin at different final concentrations of 0,20,50,100 and 200 μmol/L. After 12 hours, cell viability, immunofluorescence and RNA-sequencing(RNA-seq) were performed. Differential gene expression analysis (DEG), KEGG pathway analysis, and protein-protein interaction network analysis were conducted. Results Cisplatin decreased cell viability and increased DNA damage in HepG2,Huh7 cells. Among the genes regulated after cisplatin treatment at different concentrations, 59 genes were commonly up-regulated in both HepG2 and Huh7 cells, while 81 genes were commonly down-regulated. The commonly upregulated genes were mainly enriched in cancer initiation and progression pathways.The 81 commonly down-regulated genes were mainly enriched in Rap1 signaling pathway, Ras signaling pathway, signaling pathways regulating pluripotency of stem cells, axon guidance, and cell adhesion-related pathways. Survival analysis of key nodes in the protein-protein interaction network of commonly upregulated and downregulated genes revealed a significant correlation between high expression of Jun proto-oncogene, AP-1 transcription factor subunit (JUN) and prolonged patient survival and a significant correlation between low expression of growth arrest and DNA damage inducible alpha (GADD45A) and prolonged patient survival. Conclusions The study revealed common transcriptional changes in liver cancer cells under cisplatin treatment. Differential expression of JUN and GADD45A is a potential core mechanism to explain drug resistance. This conclusion provides some important prognostic indicators for clinical treatment.

Key words: liver cancer, transcriptome sequencing, cisplatin, gene transcription

中图分类号:

郭鑫, 冀梦蝶, 王琦, 李雪媛, 陈阳. 鉴定顺铂对人肝癌细胞系转录物组的影响[J]. 基础医学与临床, 2024, 44(3): 352-360.

GUO Xin, JI Mengdie, WANG Qi, LI Xueyuan, CHEN Yang. Identification of the effect of cisplatin on the transcriptome of human hepatocellular carcinoma cell lines[J]. Basic & Clinical Medicine, 2024, 44(3): 352-360.

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鉴定顺铂对人肝癌细胞系转录物组的影响

Identification of the effect of cisplatin on the transcriptome of human hepatocellular carcinoma cell lines

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