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PARP1通过调控POU2F2的表达促进肝细胞癌的进展研究
PARP1 promotes the progression of hepatocellular carcinoma by regulating expression of POU2F2
- 中国癌症杂志 2024年34卷第9期 页码：848-856
- 作者机构：
临汾市人民医院，山西医科大学第七临床医学院肝胆外科，山西 临汾 041000
- 作者简介：
[ "温自强（ORCID：0009-0001-1621-1692），硕士，主治医师。" ]
许其威（ORCID：0009-0001-9431-2077），本科，主任医师。
- 基金信息：临汾市软科学研究项目(2305)
DOI：10.19401/j.cnki.1007-3639.2024.09.005
中图分类号： R735.7收稿：2024-06-04，
修回：2024-09-06，
纸质出版：2024-09-30
- 稿件说明：
移动端阅览
温自强, 兰军良, 周博, 等. PARP1通过调控POU2F2的表达促进肝细胞癌的进展研究[J]. 中国癌症杂志, 2024,34(9):848-856.
Ziqiang WEN, Junliang LAN, Bo ZHOU, et al. PARP1 promotes the progression of hepatocellular carcinoma by regulating expression of POU2F2[J]. China Oncology, 2024, 34(9): 848-856.
温自强, 兰军良, 周博, 等. PARP1通过调控POU2F2的表达促进肝细胞癌的进展研究[J]. 中国癌症杂志, 2024,34(9):848-856. DOI： 10.19401/j.cnki.1007-3639.2024.09.005.
Ziqiang WEN, Junliang LAN, Bo ZHOU, et al. PARP1 promotes the progression of hepatocellular carcinoma by regulating expression of POU2F2[J]. China Oncology, 2024, 34(9): 848-856. DOI： 10.19401/j.cnki.1007-3639.2024.09.005.
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
PARP
1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，POU2类同源框2（POU class homeobox 2，
POU
2
F
2）可能是PARP1的潜在靶基因。相关性分析、RTFQ-PCR和Western blot检测一致证实POU2F2的表达受到PARP1的调控，但PJ34不能抑制POU2F2的表达。CCK-8、流式细胞术和RTFQ-PCR结果显示，采用共转染的方式在敲低
PARP
1的HCC细胞系中回补POU2F2可以增强HCC细胞增殖能力，提高HCC细胞活性，促进HCC细胞干性。
结论：
PARP1可以通过非酶活性正向调控POU2F2表达促进HCC细胞恶性表型，本研究结果有望为HCC的临床治疗和新药开发提供新的思路。
Abstract
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. Ba
sed 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 transwell assay indicated that inhibition of PARP1 attenuated proliferation and activity of HCC cells
as well as weakened their stemness
migration and invasion. Bioinformatics analysis suggested that PARP1-regulated genes were enriched in the nuclear factor-κB (NF-κB) and necroptosis pathways
with POU class homeobox 2 (
POU
2
F
2) potentially being a target gene of PARP1. Correlation analysis
along with RTFQ-PCR and Western blot detection
confirmed that the expression of POU2F2 was regulated by PARP1
while not affected by PJ34
indicating the effect of nonenzymatic function of PARP1 on POU2F2. CCK-8
flow cytometry and RTFQ-PCR results showed that the reintroduction of POU2F2 enhanced proliferative capacity
increased activity
and promoted stemness of HCC cell lines with
PARP
1 knockdown.
Conclusion:
By positively regulating the expression of POU2F2
PARP1 pr
omotes malignant phenotypes of HCC cells
providing new insights for clinical treatment and drug development for HCC.
关键词
Keywords
references
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