S100A16 silencing could inhibit the invasion and epithelial–mesenchymal transition (EMT) in GC cells. MKN45 and SGC-7901 cells were transfected with si-S100A16 or si-NC. (A) The protein levels of S100A16 were determined by Western blot. (B) Cell proliferation was detected by cell counting kit-8 (CCK-8). (C) The invasion of GC cells was assessed by Transwell assay. (D) Levels of total matrix metalloproteinase-2 (MMP-2), MMP-9, and tissue inhibitor of metalloproteinase-1 (TIMP-1) were detected in the culture supernatants of cultured GC cells by enzyme-linked immunosorbent assay (ELISA) assays. (E) The expressions of E-cadherin, N-cadherin, and vimentin were determined by qRT-PCR. All data are presented as mean ± SEM, n = 4. #p < 0.05, ##p < 0.01 versus si-NC.