The influence of extracellular matrix (ECM) stiffness on cell behavior is a well-established phenomenon. Tumor development is associated with the stiffening of the ECM. However, the understanding of the role of biomechanical behavior and mechanotransduction pathways in the oncogenesis of tumor cells remains limited. In this study, we constructed in vitro models using Polydimethylsiloxane substrates to create soft and stiff substrates. We then evaluated the migration of lung cancer cells A549 using video-microscopy and transwell assays. The mechanical properties were assessed through the utilization of atomic force microscopy, Optical Magnetic Twisting Cytometry, and traction force analysis. Additionally, the expression of Calponin 3 (CNN3) was evaluated using reverse transcription‑quantitative PCR and immunofluorescence techniques. Our observations indicate that the presence of a stiff substrate enhances A549 motility, as evidenced by increased stiffness and traction force in A549 cells on the stiff substrate. Furthermore, we observed a decrease in CNN3 expression in A549 cells on the stiff substrate. Notably, when CNN3 was overexpressed, it effectively inhibited the migration and invasion of A549 cells on the stiff substrate. The results of our study provide novel perspectives on the mechanisms underlying cancer cell migration in response to substrate mechanical properties.

中文翻译：

---

钙调蛋白 3 对肺癌细胞运动的调节是通过其感知和响应基质刚度变化的能力来实现的


细胞外基质 （ECM） 刚度对细胞行为的影响是一个公认的现象。肿瘤发展与 ECM 的硬化有关。然而，对生物力学行为和机械转导途径在肿瘤细胞肿瘤发生中的作用的理解仍然有限。在这项研究中，我们使用聚二甲基硅氧烷底物构建了体外模型，以创建柔软和坚硬的底物。然后，我们使用视频显微镜和 transwell 测定评估了肺癌细胞 A549 的迁移。通过使用原子力显微镜、光学磁扭转细胞术和牵引力分析来评估机械性能。此外，使用逆转录-定量 PCR 和免疫荧光技术评估钙调蛋白 3 （CNN3） 的表达。我们的观察表明，坚硬基质的存在增强了 A549 的运动性，刚性基质上 A549 细胞的刚度和牵引力增加证明了这一点。此外，我们观察到刚性基质上 A549 细胞中 CNN3 表达的降低。值得注意的是，当 CNN3 过表达时，它有效地抑制了 A549 细胞在坚硬基质上的迁移和侵袭。我们的研究结果为响应底物机械特性的癌细胞迁移机制提供了新的视角。