Abstract：
Growing evidence has suggested that increased matrix stiffness can significantly strengthen the malignant characteristics of hepatocellular carcinoma (HCC) cells. However, whether and how increased matrix stiffness regulates the formation of invadopodia in HCC cells remain largely unknown. In the study, we developed different experimental systems in vitro and in vivo to explore the effects of matrix stiffness on the formation of invadopodia and its relevant molecular mechanism. Our results demonstrated that increased matrix stiffness remarkably augmented the migration and invasion abilities of HCC cells, upregulated the expressions of invadopodia-associated genes and enhanced the number of invadopodia. Two regulatory pathways contribute to matrix stiffness-driven invadopodia formation together in HCC cells, including direct triggering invadopodia formation through activating integrin β1 or Piezo1/ FAK/Src/Arg/cortactin pathway, and indirect stimulating invadopodia formation through improving EGF production to activate EGFR/Src/Arg/cortactin pathway. Src was identified as the common hub molecule of two synergistic regulatory pathways. Simultaneously, activation of integrin β1/RhoA/ROCK1/MLC2 and Piezo1/Ca2+/MLCK/MLC2 pathways mediate matrix stiffness-reinforced cell migration. This study uncovers a new mechanism by which mechanosensory pathway and biochemical signal pathway synergistically regulate the formation of invadopodia in HCC cells.
摘要：
越来越多的证据表明，增加的基质硬度可以显着加强肝细胞癌（HCC）细胞的恶性特征。然而,增加的基质硬度是否以及如何调节HCC细胞内翻的形成仍然未知。在研究中,我们在体外和体内建立了不同的实验系统，以探讨基质刚度对invadopodia形成的影响及其相关分子机制。我们的结果表明，增加的基质刚度显着增强肝癌细胞的迁移和侵袭能力，上调invadopodia相关基因的表达并增加invadopodia的数量。两种调节途径有助于在HCC细胞中一起形成基质刚度驱动的invadopodia，包括通过激活整合素β1或Piezo1/FAK/Src/Arg/cortactin途径直接触发侵袭足形成，并通过改善EGF的产生来激活EGFR/Src/Arg/cortactin通路，从而间接刺激侵袭足病的形成。Src被鉴定为两个协同调节途径的共同hub分子。同时,整合素β1/RhoA/ROCK1/MLC2和Piezo1/Ca2/MLCK/MLC2途径的激活介导基质刚度增强的细胞迁移。这项研究揭示了机械感觉途径和生化信号途径协同调节HCC细胞内翻足病形成的新机制。
