摘要
目的·探讨ATP结合盒转运蛋白A7(ATP-binding cassette subfamily A member 7,ABCA7)基因在结直肠癌(colorectal cancer,CRC)中的预后评估价值,为CRC的精准诊断和个性化治疗提供新型分子标志物。方法·从肿瘤免疫单细胞数据库(Tumor Immune Single-cell Hub,TISCH)获取CRC的单细胞转录组数据,从基因表达综合(Gene Expression Omnibus,GEO)数据库下载GSM8594571样本的空间转录组数据(基于10x Genomics Visium平台),从癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库提取TCGA-COAD和TCGA-READ项目的转录组数据。通过多组学整合分析策略,利用单细胞转录组数据解析ABCA7在不同细胞亚群的表达模式,结合空间转录组数据探究其在肿瘤及邻近组织中的空间分布特征;采用比例风险回归模型(proportional hazards model,Cox模型)评估ABCA7的预后价值;通过成簇规律间隔短回文重复序列(clustered regularly interspaced short palindromic repeats,CRISPR)敲除技术结合CERES算法验证其对CRC细胞增殖的影响;利用Cistrome数据库的ChIP-seq数据及Pearson相关性分析,构建ABCA7与转录因子锌指BTB结构域蛋白33(zinc finger BTB domain-containing protein 33,ZBTB33)、核因子κB抑制因子(nuclear factor-κBrepressing factor,NKRF)的转录调控网络。结果·分析发现,ABCA7在CRC恶性细胞中显著高表达,尤其在Ki-67高表达的肿瘤组织中表达进一步上调,且肿瘤组织中ABCA7mRNA表达显著高于正常组织(P<0.05),具有一定诊断效能[受试者操作特征(receiver operating characteristic,ROC)曲线下面积(area under curve,AUC)=0.686]。敲除ABCA7可显著抑制CRC细胞系(如C75、CL11)的增殖能力。ABCA7高表达组患者的总生存期(overall survival,OS)、疾病特异性生存期(disease-specific survival,DSS)和无进展生存期(progression-free interval,PFI)均显著缩短(均P<0.05)。表观遗传分析显示,ZBTB33和NKRF可直接结合ABCA7基因调控区域,形成ZBTB33/NKRF-ABCA7转录调控轴,且三者在CRC中均显著上调表达(P<0.05)。结论·ABCA7在CRC中高表达且与不良预后相关,ZBTB33/NKRF-ABCA7调控轴可能是驱动CRC进展的关键机制。
Objective·To investigate the prognostic value of the ATP-binding cassette subfamily A member 7(ABCA7)gene in colorectal cancer(CRC)and provide a novel molecular marker for the precise diagnosis and personalized treatment of CRC.Methods·Single-cell transcriptome data of CRC were retrieved from the Tumor Immune Single-cell Hub(TISCH).Spatial transcriptome data of sample GSM8594571(based on the 10x Genomics Visium platform)were downloaded from the Gene Expression Omnibus(GEO)database.Transcriptome data from the TCGA-COAD and TCGA-READ projects were extracted from The Cancer Genome Atlas(TCGA)database.Using a multi-omics integration analysis strategy,single-cell transcriptome data were employed to analyze the expression patterns of ABCA7 in different cell subpopulations and were combined with spatial transcriptome data to explore its spatial distribution characteristics in tumor and adjacent tissues.The Cox Proportional Hazards Model(Cox)was used to evaluate the prognostic value of ABCA7.The effect of ABCA7 on CRC cell proliferation was verified using Clustered Regularly Interspaced Short Palindromic Repeats(CRISPR)knockout technology combined with the CERES algorithm.The transcriptional regulatory network between ABCA7 and the transcription factors zinc finger BTB domain-containing protein 33(ZBTB33)and Nuclear factor-κB-repressing factor(NKRF)was constructed using chromatin immunoprecipitation sequencing(ChIPseq)data from the Cistrome database and Pearson correlation analysis.Results·The analysis revealed that ABCA7 was significantly overexpressed in CRC malignant cells,with further upregulation particularly in tumor tissues with high Ki-67 expression.Moreover,the mRNA expression of ABCA7 in tumor tissues was significantly higher than that in normal tissues(P<0.05),showing certain diagnostic efficacy[area under the curve(AUC)of the receiver operating characteristic(ROC)curve=0.686].Knockout of ABCA7 significantly inhibited the proliferative capacity of CRC cell lines(e.g.,C75,CL11).Patients in the ABCA7 high-expression group had significantly shorter overall survival(OS),disease-specific survival(DSS),and progression-free interval(PFI)(all P<0.05).Epigenetic analysis showed that ZBTB33 and NKRF could directly bind to the regulatory region of the ABCA7 gene,forming a ZBTB33/NKRF-ABCA7 transcriptional regulatory axis,and all three were significantly upregulated in CRC(P<0.05).Conclusion·ABCA7 is highly expressed in CRC and associated with poor prognosis.The ZBTB33/NKRF-ABCA7 regulatory axis may be a key mechanism driving CRC progression.
作者
孙瑞状
乔坤朋
李璞
徐小莲
孟俊
SUN Ruizhuang;QIAO Kunpeng;LI Pu;XU Xiaolian;MENG Jun(Department of Laboratory Medicine,Ruijin-Hainan Hospital Shanghai Jiao Tong University School of Medicine(Hainan Boao Research Hospital),Qionghai 571437,China;Department of Pathology,Ruijin-Hainan Hospital Shanghai Jiao Tong University School of Medicine(Hainan Boao Research Hospital),Qionghai 571437,China;Department of Laboratory Medicine,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)
出处
《上海交通大学学报(医学版)》
北大核心
2025年第11期1490-1501,共12页
Journal of Shanghai Jiao tong University:Medical Science
关键词
ATP结合盒转运蛋白A7
结直肠癌
空间转录组
单细胞测序
CRISPR技术
表观遗传
转录调控轴
ATP-binding cassette subfamily A member 7(ABCA7)
colorectal cancer
spatial transcriptomics
single-cell sequencing
CRISPR technology
epigenetics
transcriptional regulatory axis
