Li Yingnan, Shi Mengjiao, Bie Beibei, Tian Hongwei, Li Jun, Li Zongfang, Sun Jin
Department of General Surgery, National-Local Joint Engineering Research Center of Biodiagnostics and Biotherapy, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710004, People's Republic of China.
Center for Tumor and Immunology, The Precision Medical Institute, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710115, People's Republic of China.
J Hepatocell Carcinoma. 2025 May 7;12:891-908. doi: 10.2147/JHC.S516656. eCollection 2025.
DDX11 antisense RNA 1 (DDX11-AS1) has been recognized for its strong correlation with hepatocellular carcinoma (HCC). Nevertheless, the exact biological functions and fundamental molecular processes of DDX11-AS1 in HCC require further in-depth investigation.
A comprehensive bioinformatics analysis was carried out to explore the expression of DDX11-AS1 and its clinical implication in HCC utilizing the TCGA data. qRT-PCR was employed to validate the expression of DDX11-AS1 in HCC tissues/cell lines. RNA fluorescence in situ hybridization (RNA-FISH) was used to observe the subcellular localization of DDX11-AS1 in HCC cells. Loss-of-function experiments, both in vitro and in vivo, were executed to elucidate the biological functions of DDX11-AS1 in HCC. RNA sequencing (RNA-seq) was employed to identify genes and signaling pathways potentially regulated by DDX11-AS1. Rescue experiments were conducted to validate that carbonic anhydrase IX (CA9) mediates DDX11-AS1 promoting HCC progression. The influence of nuclear respiratory factor 1 (NRF1) on the transcription of DDX11-AS1 was investigated through dual-luciferase reporter assays and ChIP-qPCR.
The increased expression of DDX11-AS1 is positively associated with several aggressive clinical characteristics (pathologic T stage, histologic grade, AFP level, and vascular invasion), and is closely linked to unfavorable outcomes in HCC patients, acting as a separate hazardous factor for overall survival. DDX11-AS1 is predominantly situated in the nucleus of HCC cells. DDX11-AS1 knockdown impeded the growth, migration, and invasion capabilities of HCC cells in vitro, and reduced the tumor enlargement in a subcutaneous mouse model. RNA-Seq unveiled that silencing DDX11-AS1 lessened the expression of CA9 and suppressed the activity of the MEK/ERK signaling cascade in HCC cells. Rescue experiments uncovered that CA9 acts as a downstream target facilitating the cancer-causing roles of DDX11-AS1 in HCC. Furthermore, DDX11-AS1 was revealed to be transcriptionally regulated by NRF1.
DDX11-AS1, a NRF1-induced lncRNA, facilitates HCC development by upregulating CA9 expression and activating the MEK/ERK signaling cascade.
DDX11反义RNA1（DDX11-AS1）已被证实与肝细胞癌（HCC）密切相关。然而，DDX11-AS1在HCC中的确切生物学功能和基本分子过程仍需进一步深入研究。
利用TCGA数据进行全面的生物信息学分析，以探讨DDX11-AS1在HCC中的表达及其临床意义。采用qRT-PCR验证DDX11-AS1在HCC组织/细胞系中的表达。运用RNA荧光原位杂交（RNA-FISH）观察DDX11-AS1在HCC细胞中的亚细胞定位。通过体内外功能缺失实验阐明DDX11-AS1在HCC中的生物学功能。采用RNA测序（RNA-seq）鉴定可能受DDX11-AS1调控的基因和信号通路。进行挽救实验以验证碳酸酐酶IX（CA9）介导DDX11-AS1促进HCC进展。通过双荧光素酶报告基因检测和ChIP-qPCR研究核呼吸因子1（NRF1）对DDX11-AS1转录的影响。
DDX11-AS1表达增加与多种侵袭性临床特征（病理T分期、组织学分级、甲胎蛋白水平和血管侵犯）呈正相关，且与HCC患者的不良预后密切相关，是总生存的独立危险因素。DDX11-AS1主要位于HCC细胞核中。敲低DDX11-AS1可体外抑制HCC细胞的生长、迁移和侵袭能力，并减少皮下小鼠模型中的肿瘤生长。RNA-Seq显示，沉默DDX11-AS1可降低HCC细胞中CA9的表达并抑制MEK/ERK信号级联的活性。挽救实验发现，CA9作为下游靶点促进DDX11-AS1在HCC中的致癌作用。此外，研究发现DDX11-AS1受NRF1转录调控。
DDX11-AS1是一种NRF1诱导的长链非编码RNA，通过上调CA9表达和激活MEK/ERK信号级联促进HCC发展。
