LAUSR

Cancer-associated fibroblasts (CAFs) are a key component of tumor microenvironment and are essential for tumorigenesis and development. Regorafenib is a multikinase inhibitor that targets CAFs and suppresses tumor growth. Here, we investigated the effects of regorafenib on gastrointestinal CAFs and the underlying molecular mechanisms. First, we established two in vivo tumor models, the cancer cell line HCT116 with or without mesenchymal stem cells (MSCs) and treated them with regorafenib. We found that the application of regorafenib potently impaired tumor growth, an effect that was more pronounced in tumors with a high stromal ratio, thus demonstrating that regorafenib can inhibit CAFs proliferation and induce CAFs apoptosis in vivo. Moreover, we showed that regorafenib affected macrophage infiltration by reducing the proportion of CAFs in tumors. Afterward, we induced MSCs into CAFs with exosomes to establish an in vitro model. Then, we used MTS and flow cytometry to detect the effects of regorafenib on the proliferation and apoptosis of CAFs, and Western blot to determine the expression level of apoptosis-related proteins. We found that regorafenib inhibited the proliferation of CAFs and induced the apoptosis of CAFs in vitro. Furthermore, Western blot results showed that regorafenib down-regulated the expression of B-cell lymphoma-2 (Bcl-2) and concurrently up-regulated the expression of Bcl-2-associated X (Bax), and regorafenib inhibited the phosphorylation pathway of AKT in CAFs. In conclusion, our results provide a model in which regorafenib induces CAFs apoptosis by inhibiting the phosphorylation of AKT, and regorafenib affects macrophage infiltration by reducing the proportion of CAFs in tumor tissues.