Hsu Chao-Yu, Li Jyun-Yi, Huang Wei-Ru, Liao Tsai-Ling, Wen Hsiao-Wei, Wang Chi-Young, Lye Lon-Fye, Nielsen Brent L, Liu Hung-Jen
Division of Urology, Department of Surgery, Tungs' Taichung MetroHarbor Hospital, Taichung, Taiwan.
Institute of Molecular Biology, National Chung Hsing University, Taichung, Taiwan.
Front Cell Infect Microbiol. 2025 Jun 12;15:1603124. doi: 10.3389/fcimb.2025.1603124. eCollection 2025.
Avian reovirus (ARV) is an oncolytic virus that induces autophagy and apoptosis in cancer cells, modulates the immune response, and exposes tumor-associated antigens to the immune system, making it a promising candidate for cancer therapy. Cancer cell migration and invadopodia formation are essential processes in metastasis, and targeting these mechanisms could be beneficial in limiting cancer progression.
This study investigated the effects of ARV p17 protein on cancer cell migration and invadopodia formation in HeLa and A549 cell lines. Molecular assays were conducted to examine the expression and interactions of key signaling molecules, including nucleoporin Tpr, p53, PTEN, FAK, Src, Rab40b, PI3K, Akt, TKs5, and Nck1. Analysis of TKs5, Nck1, and Rab40b mRNA levels by quantitative real-time RT-PCR. Furthermore, invadopodia detection, gelatin degradation assay, and Fluorescence imaging was performed to visualize invadopodia structures and assess extracellular matrix degradation. Additionally, rescue experiments were performed by co-transfecting cells with mutant PTEN (C124A), TKs5, or Rab40b plasmids to confirm their roles in mediating the effects of p17.
p17 suppressed nucleoporin Tpr, resulting in the activation of p53 and upregulation of PTEN. This blocked the formation of the FAK-Src complex and inhibited the Rab40b-PI3K-Akt signaling pathway. p17 also transcriptionally downregulated TKs5, Nck1, and Rab40b, thereby reducing the formation of TKs5-Nck1 and TKs5-Rab40b complexes, which are critical for invadopodia formation. Fluorescence imaging confirmed a marked reduction in invadopodia formation and matrix degradation in cells expressing p17. Restoration of invadopodia formation upon co-transfection with mutant PTEN, TKs5, or Rab40b confirmed that these molecules are key mediators of p17's inhibitory effects.
ARV p17 inhibits cancer cell migration and invadopodia formation by activating the p53-PTEN pathway and suppressing essential signaling and scaffolding complexes (FAK-Src, Rab40b-PI3K-Akt, TKs5-Nck1, and TKs5-Rab40b). These findings suggest that p17 plays a crucial anti-metastatic role and may serve as a novel therapeutic agent for targeting invasive cancer cells.
禽呼肠孤病毒（ARV）是一种溶瘤病毒，可诱导癌细胞发生自噬和凋亡，调节免疫反应，并将肿瘤相关抗原暴露于免疫系统，使其成为癌症治疗的一个有前景的候选者。癌细胞迁移和侵袭伪足形成是转移过程中的关键步骤，针对这些机制可能有助于限制癌症进展。
本研究调查了ARV p17蛋白对HeLa和A549细胞系中癌细胞迁移和侵袭伪足形成的影响。进行分子分析以检测关键信号分子的表达和相互作用，包括核孔蛋白Tpr、p53、PTEN、黏着斑激酶（FAK）、Src、Rab40b、磷脂酰肌醇-3激酶（PI3K）、蛋白激酶B（Akt）、TKs5和衔接蛋白Nck1。通过定量实时逆转录聚合酶链反应分析TKs5、Nck1和Rab40b的mRNA水平。此外，进行侵袭伪足检测、明胶降解试验和荧光成像以观察侵袭伪足结构并评估细胞外基质降解。另外，通过将细胞与突变型PTEN（C124A）、TKs5或Rab40b质粒共转染进行拯救实验，以确认它们在介导p17作用中的作用。
p17抑制核孔蛋白Tpr，导致p53激活和PTEN上调。这阻断了FAK-Src复合物的形成并抑制了Rab40b-PI3K-Akt信号通路。p17还转录下调TKs5、Nck1和Rab40b，从而减少了对侵袭伪足形成至关重要的TKs5-Nck1和TKs5-Rab40b复合物的形成。荧光成像证实表达p17的细胞中侵袭伪足形成和基质降解明显减少。与突变型PTEN、TKs5或Rab40b共转染后侵袭伪足形成的恢复证实这些分子是p17抑制作用的关键介质。
ARV p17通过激活p53-PTEN途径并抑制关键信号和支架复合物（FAK-Src、Rab40b-PI3K-Akt、TKs5-Nck1和TKs5-Rab40b）来抑制癌细胞迁移和侵袭伪足形成。这些发现表明p17发挥关键的抗转移作用，可能作为一种靶向侵袭性癌细胞的新型治疗药物。
