中华肝脏病杂志2024,Vol.32Issue(8) :734-743.DOI:10.3760/cma.j.cn501113-20240407-00176

敲低SUV3抑制肝癌细胞增殖、迁移和侵袭并诱导细胞程序性死亡受体配体1表达

SUV3 knockdown inhibits proliferation,migration,and invasion of hepatocellular carcinoma cells and induces PD-L1 expression

张俊昊 吴骁 汪婷婷 汪子书
中华肝脏病杂志2024,Vol.32Issue(8) :734-743.DOI:10.3760/cma.j.cn501113-20240407-00176

敲低SUV3抑制肝癌细胞增殖、迁移和侵袭并诱导细胞程序性死亡受体配体1表达

SUV3 knockdown inhibits proliferation,migration,and invasion of hepatocellular carcinoma cells and induces PD-L1 expression

张俊昊 1吴骁 1汪婷婷 1汪子书1
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作者信息

  • 1. 蚌埠医科大学附属第一医院肿瘤内科,安徽 233004
  • 折叠

摘要

目的 研究SUV3基因在肝癌发生和发展过程中的作用.方法 通过分析TCGA和GTEx数据库中的转录组测序数据比较SUV3在肝癌组织和正常肝组织中的表达差异.用RNA干扰技术在不同肝癌细胞中敲低SUV3,以及构建过表达载体在不同的肝癌细胞中过表达SUV3,研究SUV3对肝癌细胞增殖迁移侵袭的调节作用.亚细胞组分分离实验用于研究敲低SUV3是否会导致线粒体DNA释放到细胞质中.用定量逆转录PCR技术研究敲低SUV3是否影响细胞程序性死亡受体配体1(PD-L1)的表达.两组间比较应用双侧t检验.结果 分析TCGA数据库结果显示,SUV3在肝癌组织中的表达高于正常肝组织,肝癌组织中SUV3表达高的肝癌患者预后不良,定量RT-PCR结果表明SUV3在肝癌组织中的表达高于癌旁肝组织.MTS实验结果显示SUV3敲低的肝癌细胞增殖速率明显低于对照肝癌细胞(P<0.01),SUV3过表达的肝癌细胞增殖速率明显高于对照肝癌细胞(P<0.01).细胞划痕实验和细胞迁移、侵袭实验结果显示敲低SUV3抑制肝癌细胞的迁移和侵袭(P<0.01),过表达SUV3促进肝癌细胞的迁移和侵袭(P<0.05).敲低SUV3导致线粒体DNA总体水平降低(P<0.01)的同时伴随着细胞质中线粒体DNA水平升高(P<0.01),提示敲低SUV3导致线粒体DNA泄漏到细胞质中.敲低SUV3导致PD-L1表达升高(P<0.001),SUV3敲低的细胞中过表达TREX1降低细胞质中线粒体DNA水平,且能够抑制敲低SUV3导致PD-L1表达升高,说明敲低SUV3通过增加细胞质DNA水平诱导PD-L1表达.结论 SUV3基因可能在肝癌细胞中发挥癌基因的功能.

Abstract

Objective To study the SUV3 gene role during the process of occurrence and advancement of hepatocellular carcinom.Methods The The differences in SUV3 expression between hepatocellular carcinoma tissues and normal liver tissues were compared by analyzing transcriptome sequencing data from TCGA and GTEx databases.SUV3 knockdown in different hepatocellular carcinoma cells was performed using RNA interference technology.Overexpression vectors were constructed to overexpress SUV3 in different hepatocellular carcinoma cells.The SUV3 regulatory effect was studied on proliferation,migration,and invasion of hepatocellular carcinoma cells.A subcellular fraction isolation approach was used to investigate whether SUV3 knockdown resulted in the release of mitochondrial DNA into the cytoplasm.Quantitative reverse transcription PCR was applied to investigate whether SUV3 knockdown affected PD-L1 expression.The two groups were compared using a two-tailed t-test.Results The TCGA database analysis revealed that SUV3 expression was higher in hepatocellular carcinoma tissues than in normal liver tissues,and the prognosis of patients with high SUV3 expression in hepatocellular carcinoma tissues was poor.The quantitative RT-PCR results showed that SUV3 expression was higher in hepatocellular carcinoma tissues than that in paracancerous liver tissue.The MTS assay showed that with SUV3 knockdown,the proliferation rate was significantly lower in hepatocellular carcinoma cells than that of the control hepatocellular carcinoma cells(P<0.01).The proliferation rate was significantly higher in SUV3-overexpressed hepatocellular carcinoma cells than that of control hepatocellular carcinoma cells(P<0.01).Cell scratch assay and cell migration and invasion assay showed that SUV3 knockdown inhibited the migration and invasion of hepatocellular carcinoma cells(P<0.01),while SUV3 overexpression promoted the migration and invasion of hepatocellular carcinoma cells(P<0.05).SUV3 Knockdown led to a decrease in the overall level of mtDNA(P<0.01)in accompanied by an increase in mtDNA level in the cytoplasm(P<0.01),indicating that SUV3 knockdown led to mitochondrial DNA leakage into the cytoplasm.SUV3 knockdown resulted in elevated PD-L1 expression(P<0.001),and overexpression of TREX1 in SUV3 knockdown cells decreased mtDNA levels in the cytoplasm and inhibited SUV3 knockdown,resulting in elevated PD-L1 expression,indicating that SUV3 knockdown induced PD-L1 expression by increasing cytoplasmic DNA levels.Conclusions The SUV3 gene may play an oncogenic function in hepatocellular carcinoma cells.

关键词

肝细胞癌/细胞/增殖/迁移/侵袭/程序性死亡受体配体1

Key words

Hepatocellular carcinoma/Cell/Proliferation/Migration/Infestation/Programmed death-ligand 1

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基金项目

安徽省临床医学研究转化专项项目(202204295107020038)

出版年

2024
中华肝脏病杂志
中华医学会

中华肝脏病杂志

CSTPCDCSCD北大核心
影响因子:1.625
ISSN:1007-3418
参考文献量29
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