Liang Libo, Wang Xinyi, Zeng Yuping, Chen Hao, Zhou Wen, Mu Hongying, Liao G A
General Practice Medical Center, West China Hospital, Sichuan University, Chengdu, 610041, China.
Department of Laboratory Medicine, West China Hospital, Sichuan University, Chengdu, 610041, China.
Oncol Res. 2025 May 29;33(6):1405-1421. doi: 10.32604/or.2024.056708. eCollection 2025.
Exosomal long noncoding RNAs (lncRNAs) might facilitate epithelial-mesenchymal transition (EMT) in liver cancer after transarterial chemoembolization (TACE), thereby enhancing tumor cell invasiveness and migration. This study investigated the prognostic role of plasma exosomal long noncoding RNA-plasmacytoma variant translocation 1 (lncRNA-PVT1) in TACE treated hepatocellular carcinoma (HCC).
Plasma exosomal lncRNA-PVT1 was evaluated via qPCR before and after TACE. Hepatoma cell behavior was investigated in different HCC cell lines. A lncRNA-PVT1 plasmid was synthesized and overexpressed, and si-lncRNA PVT1 was transfected into poorly invasive cells to reveal its influence on cell characteristics. The lncRNA-PVT1-FoxM1 interaction was elucidated using a double-luciferase reporter gene assay. The effect of miRNA-345-5p on minimally invasive hepatoma cells was assessed. Three experimental groups were established: poorly invasive cells, poorly invasive cells co-cultured with exosomes from hypoxia-induced highly invasive cells, and poorly invasive cells co-cultured with normal hepatocyte exosomes. Liver cancer cells were subcutaneously inoculated into nude mice, with subsequent observations of weight, tumor formation, and tumor size.
We identified two lncRNAs (lncRNA-PVT1 and GAPLINC) associated with EMT in the hypoxic microenvironment of liver cancer. Cox multivariate regression analysis was used to establish a prognostic model distinguishing high- and low-risk groups. Hypoxia-induced HepG2 exosomes significantly promoted EMT in poorly invasive HCC cells. LncRNA-PVT1 overexpression and silencing altered E-cadherin, vimentin, and FoxM1 expression, cell proliferation, invasion, migration, and apoptosis. miR-345-5p directly targeted lncRNA-PVT1 and FoxM1, affecting downstream targets. , co-culturing poorly invasive hepatoma cells with exosomes from highly invasive cells increased tumor volumes, upregulated lncRNA-PVT1, FoxM1, Ki67, and MMP9 expression, and downregulated miR-345-5p expression.
Plasma exosomal lncRNA-PVT1 expression is upregulated in highly invasive cells post-hypoxia, potentially serving as a biomarker for evaluating liver cancer prognosis after TACE. Through a miRNA-345-5p-mediated competing endogenous RNA mechanism, it promotes EMT in poorly invasive cells, likely contributing to recurrence and metastasis post-HCC interventional embolization.
外泌体长链非编码RNA（lncRNAs）可能在经动脉化疗栓塞术（TACE）后促进肝癌上皮-间质转化（EMT），从而增强肿瘤细胞的侵袭性和迁移能力。本研究调查了血浆外泌体长链非编码RNA-浆细胞瘤变异易位1（lncRNA-PVT1）在TACE治疗的肝细胞癌（HCC）中的预后作用。
通过qPCR评估TACE前后血浆外泌体lncRNA-PVT1水平。在不同的HCC细胞系中研究肝癌细胞行为。合成并过表达lncRNA-PVT1质粒，将si-lncRNA PVT1转染至低侵袭性细胞中，以揭示其对细胞特性的影响。使用双荧光素酶报告基因检测法阐明lncRNA-PVT1与FoxM1的相互作用。评估miRNA-345-5p对低侵袭性肝癌细胞的作用。建立三个实验组：低侵袭性细胞、与缺氧诱导的高侵袭性细胞来源的外泌体共培养的低侵袭性细胞、与正常肝细胞来源的外泌体共培养的低侵袭性细胞。将肝癌细胞皮下接种到裸鼠体内，随后观察体重、肿瘤形成和肿瘤大小。
我们在肝癌缺氧微环境中鉴定出两种与EMT相关的lncRNAs（lncRNA-PVT1和GAPLINC）。采用Cox多因素回归分析建立区分高风险和低风险组的预后模型。缺氧诱导的HepG2外泌体显著促进低侵袭性HCC细胞的EMT。lncRNA-PVT1的过表达和沉默改变了E-钙黏蛋白、波形蛋白和FoxM1的表达、细胞增殖、侵袭、迁移和凋亡。miR-345-5p直接靶向lncRNA-PVT1和FoxM1，影响下游靶点。将低侵袭性肝癌细胞与高侵袭性细胞来源的外泌体共培养可增加肿瘤体积，上调lncRNA-PVT1、FoxM1、Ki67和MMP9的表达，并下调miR-345-5p的表达。
缺氧后高侵袭性细胞中血浆外泌体lncRNA-PVT1表达上调，可能作为评估TACE后肝癌预后的生物标志物。通过miRNA-345-5p介导的竞争性内源RNA机制，它促进低侵袭性细胞的EMT，可能导致HCC介入栓塞后复发和转移。
