摘要
背景与目的:肝细胞癌(hepatocellular carcinoma,HCC)是严重威胁人类健康的重大疾病。多腺苷二磷酸核糖聚合酶1[poly(ADP-ribose)polymerase 1,PARP1]作为一种腺苷二磷酸核糖转移酶,能够促进DNA损伤修复进程,因此,PARP1通常被看作是一种促癌基因,但其在HCC中的表达和机制尚不明确。本研究旨在探讨PARP1在HCC患者中的表达趋势及其在HCC发生、发展中的作用。方法:首先,通过对癌症基因组图谱(The Cancer Genome Atlas,TCGA)和临床蛋白质组学癌症分析联盟(Clinical Proteomic Tumor Analysis Consortium,CPTAC)HCC数据库的分析鉴定PARP1的临床表达情况,采用实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)和蛋白质印迹法(Western blot)检测PARP1在HCC患者癌组织及癌旁组织样本中的表达情况。然后,借助PARP1的抑制剂PJ34抑制PARP1酶活性,同时借助小RNA干扰技术下调HCC细胞系中PARP1的表达,并以此为模型,采用细胞计数试剂盒-8(cell counting kit-8,CCK-8)和流式细胞术检测PARP1对细胞活力的影响,采用RTFQ-PCR检测HCC细胞干性基因的表达变化,采用细胞迁移和侵袭实验检测HCC细胞迁移和侵袭能力。采用生物信息学方法分析HCC进展中受PARP1调控的靶基因及其参与通路,并通过回补实验确定PARP1靶基因是否参与HCC细胞恶性表型。结果:在TCGA和CPTAC数据库中,PAPR1的表达均在HCC组中显著上调。RTFQ-PCR和Western blot检测结果表明,相比癌旁组织,HCC组织中的PARP1在转录和翻译水平均显著上调。生存分析结果表明,PARP1的表达与HCC患者的预后呈显著负相关。CCK-8、流式细胞术、RTFQ-PCR、细胞迁移及侵袭实验结果显示,在HCC细胞中下调PARP1表达可以抑制HCC细胞增殖,降低HCC细胞活性及干性,并减弱HCC细胞迁移和侵袭能力。生物信息学分析提示,PARP1调控基因富集在核因子κB(nuclear factor-κB,NF-κB)和Necroptosis通路中,PO
Background and purpose:Hepatocellular carcinoma(HCC)is a major disease seriously threatening human health.Poly(ADP-ribose)polymerase-1(PARP1)is an enzyme that catalyzes poly ADP-ribosylation.Given the role of PARP1 in DNA damage repair,it is generally considered as an oncogene.However,the expression of PARP1 and its mechanism in HCC are not yet clear.This study aimed to investigate the role of PARP1 in the occurrence and development of HCC and its potential mechanisms.Methods:First,we analyzed the expression pattern of PARP1 in The Cancer Genome Atlas(TCGA)and Clinical Proteomic Tumor Analysis Consortium(CPTAC)HCC database,and identified the expression trend of PARP1 in our HCC cohort using real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot.Then,the enzyme activity of PARP1 was inhibited by PJ34,an inhibitor of PARP1 and the expression of PARP1 in HCC cell lines was downregulated with small RNA interference technology.Based on these models,the following experiments were conducted:First,the effect of PARP1 on cell viability was assessed by cell counting kit-8(CCK-8)assay and flow cytometry;Second,the expression levels of stemness-related genes in HCC cells were identified using RTFQ-PCR;Third,the effect of inhibition of PARP1 on migration and invasion of HCC cells was detected by migration and invasion assay(transwell assay).Finally,bioinformatic analysis was performed to identify new target genes and the pathways regulated by PARP1 in HCC progression.Rescue experiments were performed to determine whether PARP1 target genes were involved in the malignant phenotypes of HCC cells.Results:The expression of PARP1 was significantly up-regulated in HCC tissues in both TCGA and CPTAC database.RTFQ-PCR and Western blot assays showed that PARP1 was obviously up-regulated in HCC tissues compared to paracancerous tissues.Survival analysis showed that PARP1 expression was significantly negatively correlated with the prognosis of patients.The results of CCK-8,flow cytometry,RTFQ-PCR and tra
作者
温自强
兰军良
周博
许其威
WEN Ziqiang;LAN Junliang;ZHOU Bo;XU Qiwei(Department of Hepatobiliary Surgery,Linfen People’s Hospital,The Seventh Clinical Medical College of Shanxi Medical University,Linfen 041000,Shanxi Province,China)
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2024年第9期848-856,共9页
China Oncology
基金
临汾市软科学研究项目(2305)。
