Research On The Mechanism Of Legumin Promoting Tumor Metastasis Through Tumor-associated Macrophages

Objective: Tumor-associated macrophages(TAMs)play a very important role in the tumor microenvironment.Legumin is highly expressed in gastric cancer and has a close relationship with the tumor microenvironment.Does legumin regulate TAMs? The influence on the occurrence and development of gastric cancer is not clear.The purpose of this study is to explore the effect of legumin on the proliferation and migration of TAMs,and to further study whether legumin can affect the occurrence and development of gastric cancer through TAMs.Methods: Selectively activated TAMs were obtained by in vitro induction with PMA / IL-4 / IL-13;stable transfected cell lines with overexpression and knockdown of legumain were constructed using lentivirus-mediated gene overexpression or knockdown techniques;Western blot was used to detect the protein expression of legumain;CCK-8 and Transwell assays were used to detect cell proliferation and migration ability;subcutaneous tumor formation experiments in nude mice were used to verify the effect of legumain-deficient TAMs on gastric cancer at the animal level;application Immunofluorescence double staining and immunohistochemistry were used to detect the expression of CD68,legumin,CD31 and Ki67 in tumor tissues,respectively.Results: The lentiviral-mediated technology was used to successfully construct the THP-1 and Raw264.7 cell lines with overexpression and knockdown of legumain;overexpression of legumain enhanced the cell proliferation and migration ability of TAMs,whereas knockdown of legumain reduced TAMs In addition,legumin affects the proliferation and migration of gastric cancer cells in vitro and the ability to form tumors in vivo through TAMs.Conclusion:Studies have shown that legumain can regulate the proliferation and migration of TAMs cells,and that TAMs legumain can regulate the proliferation and invasion of gastric cancer cells in vitro,affecting tumor in vitro tumorigenicity and tumor angiogenesis.These results indicate that legumin plays a very important role in the regulation of gastric cancer cell proliferation and invasion by TAMs,and may serve as a new target for gastric cancer treatment.