基于转录组测序技术分析补骨脂素对HepG2细胞的作用机制*

doi:10.3969/j.issn.1671-2587.2024.01.006

临床输血与检验 ›› 2024, Vol. 26 ›› Issue (1): 35-41.DOI: 10.3969/j.issn.1671-2587.2024.01.006

基于转录组测序技术分析补骨脂素对HepG2细胞的作用机制^*

汪思云, 李艺博, 谭春霞, 卢涛

北京中医药大学生命科学学院,北京 102488

收稿日期:2023-11-14 发布日期:2024-03-13
通讯作者: 卢涛,主要从事中医药与抗衰研究,（E-mail）taolu@bucm.edu.cn。
作者简介:汪思云,主要从事中医药与抗衰研究,（E-mail）wangsiyun0710@163.com。
基金资助:
*本课题受北京市高精尖学科经费-中医生命科学（No.1000062520573）资助

Exploring the Mechanism of Psoralen on HepG2 Cells Based on Transcriptome Sequencing

WANG Siyun, LI Yibo, TAN Chunxia, LU Tao

School of Life Science,Beijing University of Chinese Medicine,Beijing 102488

Received:2023-11-14 Published:2024-03-13

摘要/Abstract

摘要： 目的利用转录组测序（RNA-sequencing,RNA-seq）技术分析补骨脂素对肝癌HepG2细胞的作用机制。方法利用CCK-8法检测不同浓度的补骨脂素（0、62.5、125、250、500 μM）作用HepG2细胞12 h和24 h后存活率的变化;流式细胞术检测250 μM补骨脂素对HepG2细胞处理24 h后细胞周期的变化;RNA-seq测序获得对照组和补骨脂素组差异表达基因并进行GO富集分析和KEGG通路分析;实时荧光定量PCR技术验证差异基因表达水平。结果与对照组相比,补骨脂素作用HepG2细胞后,其存活率明显下降（P<0.01）,细胞周期在G0/G1期比例提高（P<0.05）;RNA-seq测序分析共有286个差异显著基因,其中包括130个上调基因和156个下调基因;GO功能富集和KEGG信号通路主要集中在细胞周期相关生物学过程及信号通路;qPCR结果显示HepG2细胞经补骨脂素处理后p53和GADD45B基因上调（P<0.05,P<0.01）,CDK2、RRM2和CCND1基因下调（P<0.05,P<0.01）。结论补骨脂素对HepG2细胞产生毒性,阻滞细胞周期进程,其作用机制可能与调控细胞周期相关基因（p53,CDK2,GADD45B,CCND1,RRM2）的表达有关。

关键词: 补骨脂素, HepG2, RNA测序, 细胞周期

Abstract: Objective To investigate the effects of psoralen on HepG2 cells by transcriptome sequencing technology（RNA-seq）. Methods The effects of various concentrations of psoralen (0、62.5、125、250、500 μM) on the viability of HepG2 cells after 12 h and 24 h treatment were detected by CCK-8 assay.The cell cycle of HepG2 cells exposed to 250 μM psoralen for 24 h was detected by flow cytometry.RNA-seq followed by GO enrichment analysis and KEGG pathway enrichment analysis identified differentially expressed genes between control and psoralen groups.qPCR was conducted to verify the gene expression levels of target genes. Results Compared with the control group,the cell viability rate of HepG2 cells after treated with psoralen was significantly decreased (P<0.01), and the cell cycle in the G0/G1 phase was increased (P<0.05).There are 286 differentially expressed genes in psoralen groups,of which 130 were up-regulated and 156 were down-regulated. The GO enrichment analysis and KEGG pathway enrichment analysis were mainly involved in the biological processes and signal pathways related to the cell cycle. The qPCR results showed that p53 and GADD45B gene expression was up-regulated (P<0.05, P<0.01), CDK2, RRM2 and CCND1 gene expression was down-regulated by treatment of Psoralen in HepG2 cell (P<0.05, P<0.01). Conclusion Psoralen has toxic effects on HepG2 cells and inhibition of cell cycle progression.The mechanism may be related to the regulation of the expression of cell cycle genes (p53, CDK2, GADD45B, CCND1, RRM2).

Key words: Psoralen, HepG2, RNA sequencing, Cell cycle

中图分类号:

R285Q78

汪思云, 李艺博, 谭春霞, 卢涛. 基于转录组测序技术分析补骨脂素对HepG2细胞的作用机制^*[J]. 临床输血与检验, 2024, 26(1): 35-41.

WANG Siyun, LI Yibo, TAN Chunxia, LU Tao. Exploring the Mechanism of Psoralen on HepG2 Cells Based on Transcriptome Sequencing[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2024, 26(1): 35-41.

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基于转录组测序技术分析补骨脂素对HepG2细胞的作用机制^*

Exploring the Mechanism of Psoralen on HepG2 Cells Based on Transcriptome Sequencing

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