李丹秀¹;何阳菘¹;程浩¹;庞茂桂¹;叶小妮²;安广洲¹;周云¹;卢瑗瑗²;赵晓迪²;王新¹

• 1:空军军医大学唐都医院消化内科，陕西 西安 710038
• 2:空军军医大学西京消化病医院肿瘤生物学国家重点实验室，消化疾病国家临床研究中心，陕西 西安 710032

摘要(Abstract)：

目的 通过癌症基因组图谱（The Cancer Genome Atlas，TCGA）数据库筛选肝细胞癌（hepatocellular carcinoma，HCC）中关键的增强子RNA（enhancer RNA，eRNA），探究其作为HCC预后分子和治疗靶点的潜力。方法 从TCGA数据库中检索下载33种癌症类型的基因表达RNA-seq配置文件，应用Kaplan-Meier生存分析和Spearman相关分析筛选与HCC患者生存相关的eRNA和靶基因。评估目的eRNA与临床特征的相关性。进行基因集富集分析以研究目的eRNA潜在的生物学功能。在HCC细胞株Hep 3B中分别构建AP003469.2和靶基因酪氨酸/色氨酸羟化酶激活蛋白ζ（tyrosine and tryptophan hydroxylaseactivators zeta，YWHAZ）功能缺失细胞模型，定量实时PCR检测AP003469.2敲低后对YWHAZ表达的影响。应用体外增殖实验和transwell实验探究AP003469.2和YWHAZ敲低对细胞增殖和迁移、侵袭能力的影响。在泛癌中验证目的eRNA在其他癌种中的预后潜能。结果 在HCC中，AP003469.2和靶基因YWHAZ与患者不良预后相关，且AP003469.2的表达在患者的组织病理学分级、AJCC分期和T分期方面，差异均具有统计学意义（均P<0.05）。基因和通路富集分析结果显示，AP003469.2与参与中性粒细胞脱颗粒、中性粒细胞活化、细胞外基质降解、细胞骨架蛋白重构、细胞伪足和细胞黏附等生物学行为的分子和调控通路相关；进一步分析发现，AP003469.2可能通过中性粒细胞细胞陷阱和细胞伪足调控HCC的发生和发展。定量实时PCR实验证实AP003469.2对靶基因YWHAZ转录的促进作用（P<0.05）。CCK-8和transwell实验表明，AP003469.2和YWHAZ敲低均可抑制HCC细胞株Hep 3B的增殖、迁移和侵袭能力（均P<0.01）。结论 AP003469.2可能是HCC中的关键eRNA，有潜力成为HCC早期诊断和预后的标志物和治疗靶点。

关键词(KeyWords)： 肝细胞癌;增强子 RNA;AP003469.2;生物信息学

Abstract:

Keywords:

基金项目(Foundation):

作者(Authors): 李丹秀¹;何阳菘¹;程浩¹;庞茂桂¹;叶小妮²;安广洲¹;周云¹;卢瑗瑗²;赵晓迪²;王新¹

DOI: 10.13267/j.cnki.syzlzz.2022.022

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