p130Cas induces bone invasion by oral squamous cell carcinoma by regulating tumor epithelial-mesenchymal transition and cell proliferation

Bone invasion is a critical factor in determining the prognosis of oral squamous cell carcinoma (OSCC) patients. Transforming growth factor beta (TGF-beta) is abundantly expressed in the bone matrix and is involved in the acquisition of aggressiveness by tumors. TGF-beta) is also important to cytoskeletal changes during tumor progression. In this study, we examined the relationship between TGF-beta signaling and cytoskeletal changes during bone invasion by OSCC. Immunohistochemical staining of OSCC samples from 5 patients showed the expression of p130Cas (Crk-associated substrate) in the cytoplasm and phosphorylated Smad3 expression in the nucleus in OSCC cells. TGF-beta1 induced the phosphorylation of Smad3 and p130Cas as well as epithelial-mesenchymal transition (EMT) accompanied by the downregulation of the expression of E-cadherin, a marker of epithelial cells, and the upregulation of the expression of N-cadherin, or Snail, a marker of mesenchymal cells, in human HSC-2 cells and mouse SCCVII cells. SB431542, a specific inhibitor of Smad2/3 signaling, abrogated the TGF-beta1-induced phosphorylation of p130Cas and morphological changes. Silencing p130Cas using an shRNA or siRNA in SCCVII cells suppressed TGF-beta1-induced cell migration, invasion, EMT, and matrix metalloproteinase-9 (MMP-9) production. Compared with control SCCVII cells, SCCVII cells with silenced p130Cas strongly suppressed zygomatic and mandibular destruction in vivo by reducing the number of osteoclasts, cell proliferation and MMP-9 production. Taken together, these results showed the expression of TGF-beta/p130Cas might be a new target for the treatment of OSCC bone invasion (233 words).