Gu Xiaoyuan, Lu Hongmin, Wang Wei, Zhao Zijun, Zhang Weiqiang, Lu Xinyuan
Department of Oncology, Shibei Hospital of Shanghai, Jing'an District, No. 4500, Gonghexin Road, Shanghai, China.
Department of Oncology, Renji Hospital, School of Medicine, Shanghai Jiao Tong University, No. 145, Shandong Middle Road, Shanghai, 200127, China.
Discov Oncol. 2025 Apr 17;16(1):546. doi: 10.1007/s12672-025-02296-7.
MicroRNAs (miRNAs), particularly miR-130a-5p, play pivotal roles in the tumorigenesis and progression of hepatocellular carcinoma (HCC) by participating in diverse biological processes. The objective of this study was to elucidate the mechanistic basis by which miR-130a-5p regulates the expression of tissue factor pathway inhibitor-2 (TFPI2) and to demonstrate the subsequent impact of the miR-130a-5p/TFPI2 axis on HCC invasion.
Expression levels of miR-130a-5p and TFPI2 were quantified in HepG2 cell lines using quantitative real-time PCR (qRT-PCR). Western blot and qRT-PCR were employed to assess the expression of TFPI2 and epithelial-to-mesenchymal transition (EMT)-related proteins in both cancer cells and tissues. miR-130a-5p knockdown and TFPI2 overexpression were achieved through transfection of HepG2 cells with short hairpin RNA (shRNA) and synthetic overexpression plasmids, respectively. A dual luciferase reporter assay was conducted to verify the binding of miR-130a-5p to TFPI2. Migration and invasion capabilities of cancer cells were evaluated using Transwell migration and invasion assays. A mouse xenograft tumor model was established to investigate tumor growth in vivo. Immunohistochemical (IHC) staining was utilized to examine the expression of EMT-related proteins in tumor tissues.
The dual-luciferase reporter assay confirmed that miR-130a-5p binds to the 3' untranslated region (3'UTR) of TFPI2 mRNA, inhibiting its luciferase activity. Western blot analysis revealed that miR-130a-5p negatively regulates TFPI2 protein expression and promotes EMT molecular events by targeting TFPI2 in HCC cells. Transwell assays demonstrated that downregulation of miR-130a-5p and upregulation of TFPI2 inhibited the migration and invasion abilities of HCC cells in vitro. Silencing of miR-130a-5p was found to retard the growth of HCC xenografts in vivo, decrease TFPI2 expression, and alter the EMT process.
miR-130a-5p binds to TFPI2 mRNA and promotes HCC cell migration, invasion, and xenograft tumor growth by regulating the EMT process. These findings suggest that the miR-130a-5p/TFPI2 axis may represent a promising therapeutic target for the treatment of HCC.
微小RNA（miRNA），尤其是miR-130a-5p，通过参与多种生物学过程在肝细胞癌（HCC）的肿瘤发生和进展中起关键作用。本研究的目的是阐明miR-130a-5p调节组织因子途径抑制剂2（TFPI2）表达的机制基础，并证明miR-130a-5p/TFPI2轴对HCC侵袭的后续影响。
使用定量实时PCR（qRT-PCR）在HepG2细胞系中定量miR-130a-5p和TFPI2的表达水平。采用蛋白质免疫印迹法（Western blot）和qRT-PCR评估癌细胞和组织中TFPI2及上皮-间质转化（EMT）相关蛋白的表达。分别通过用短发夹RNA（shRNA）转染HepG2细胞和合成过表达质粒实现miR-130a-5p敲低和TFPI2过表达。进行双荧光素酶报告基因检测以验证miR-130a-5p与TFPI2的结合。使用Transwell迁移和侵袭实验评估癌细胞的迁移和侵袭能力。建立小鼠异种移植肿瘤模型以研究体内肿瘤生长。利用免疫组织化学（IHC）染色检测肿瘤组织中EMT相关蛋白的表达。
双荧光素酶报告基因检测证实miR-130a-5p与TFPI2 mRNA的3'非翻译区（3'UTR）结合，抑制其荧光素酶活性。蛋白质免疫印迹分析显示，miR-130a-5p通过靶向HCC细胞中的TFPI2负调节TFPI2蛋白表达并促进EMT分子事件。Transwell实验表明，miR-130a-5p下调和TFPI2上调抑制了HCC细胞在体外的迁移和侵袭能力。发现miR-130a-5p沉默可延缓体内HCC异种移植瘤的生长，降低TFPI2表达，并改变EMT过程。
miR-130a-5p与TFPI2 mRNA结合，通过调节EMT过程促进HCC细胞迁移、侵袭和异种移植瘤生长。这些发现表明，miR-130a-5p/TFPI2轴可能是治疗HCC的一个有前景的治疗靶点。
