Buglione Arianna, Alloisio Giulia, Ciaccio Chiara, Rodriguez David Becerril, Dogali Simone, Luce Marco, Marini Stefano, Cricenti Antonio, Gioia Magda
Department of Clinical Sciences and Translational Medicine, University of Rome Tor Vergata, Via Montpellier 1, Roma 00133, Italy.
Institute of Structure Matter del Consiglio Nazionale delle Ricerche ISM-CNR, Via del Fosso del Cavaliere 100, Rome I-00133, Italy.
Eur J Cell Biol. 2025 Mar;104(1):151469. doi: 10.1016/j.ejcb.2024.151469. Epub 2024 Dec 11.
Despite their genetic diversity, metastatic cells converge on similar physical constraints during tumor progression. At the nanoscale, these forces can induce substantial molecular deformations, altering the structure and behavior of cancer cells. To address the challenges of osteosarcoma (OS), a highly aggressive cancer, we explored the mechanobiology of OS cells, in vitro. Using uniaxial-stretching technology, we examined the biophysical modulation of metastatic traits in SAOS-2, U-2 OS, and non-tumorigenic hFOB cells. Changes in cell morphology were quantified using confocal and fluorescence microscopy. To elucidate the molecular mechanisms that translate biomechanical alterations into biochemical responses, we employed Western blotting, real-time quantitative RT-PCR, reactive oxygen species ROS assay, and the mechanosensitive channel blocker Grammostola MechanoToxin4 (GsMTx-4). Our study reveals that mechanical stimulation uniquely affects OS cells, increasing nuclear size and altering the N/C ratio. We found that mechanosensitive (MS) channels are activated, leading to ROS accumulation, Src protein modulation, and histone H3 acetylation. These changes influence OS cell motility and adhesion but not proliferation. Importantly, mechanical preconditioning differentially impacts doxorubicin resistance, correlating with the Src-H3 acetylation axis. This study underscores the critical role of MS channels in OS cells and highlights the importance of mechanobiology in identifying molecular pathways that traditional biochemical approaches may not reveal. Notably, the GsMTx-4 venom peptide effectively countered mechanically induced responses, particularly by inhibiting OS cell migration, without harming healthy cells. Thus, suggesting its potential as a promising therapeutic agent for targeting osteosarcoma metastasis.
尽管转移细胞具有遗传多样性，但在肿瘤进展过程中，它们会面临相似的物理限制。在纳米尺度上，这些力可引起显著的分子变形，改变癌细胞的结构和行为。为应对骨肉瘤（OS）这一极具侵袭性的癌症所带来的挑战，我们在体外探索了OS细胞的力学生物学。利用单轴拉伸技术，我们研究了SAOS-2、U-2 OS和非致瘤性hFOB细胞中转移特征的生物物理调节。使用共聚焦显微镜和荧光显微镜对细胞形态变化进行定量分析。为阐明将生物力学改变转化为生化反应的分子机制，我们采用了蛋白质免疫印迹法、实时定量逆转录-聚合酶链反应、活性氧（ROS）检测以及机械敏感通道阻滞剂Grammostola机械毒素4（GsMTx-4）。我们的研究表明，机械刺激对OS细胞具有独特影响，可增加细胞核大小并改变核质比。我们发现机械敏感（MS）通道被激活，导致ROS积累、Src蛋白调节以及组蛋白H3乙酰化。这些变化影响OS细胞的运动性和黏附性，但不影响其增殖。重要的是，机械预处理对阿霉素耐药性有不同影响，这与Src-H3乙酰化轴相关。本研究强调了MS通道在OS细胞中的关键作用，并突出了力学生物学在识别传统生化方法可能无法揭示的分子途径方面的重要性。值得注意的是，GsMTx-4毒液肽有效对抗了机械诱导的反应，特别是通过抑制OS细胞迁移，而不损害健康细胞。因此，表明其作为一种有前景的靶向骨肉瘤转移治疗药物的潜力。
