Fig. 2

CALCR promotes tumorigenesis by regulating cell proliferation and migration. (A) Reduced CALCR protein expression in AGS and SGC-7901 cells in the shCALCR group. (B) Knockdown of CALCR inhibits proliferation in both AGS and SGC-7901 cell lines. (C) In vivo imaging system showing tumorigenesis in negative control (NC) mice and CALCR knockout (KD) mice. (D) Total and average radiation efficiency of NC mice (#1 to #10) and KD mice (#11 to #20). (E) Tumor volume and tumor mass over time in KD mice and NC mice. (F) Representative immunohistochemical staining images and relative expression levels of ki-67 in KD and NC mouse groups. Experiments were repeated at least three times. Asterisks (*) indicate a significant difference compared with control groups (*P < 0.05, **P < 0.01 and ***P < 0.001). NC, negative control. KD, knockdown. (G, H) Fluorescence activated cell sorting (FACS) showed the difference of cell apoptosis percentage between different groups. (I, J) transwell assay showed the difference of migration rate between different groups. (K) Western blot analysis showed the level of CALCR and AKT pathway-associated protein in MGC-803 cell line from shCALCR and AKT activator groups. (L) Association of CALCR with apoptosis-related gene expression. (M) Changes in expression of apoptotic genes associated with the CALCR gene as shown by Western Blot. Experiments were repeated at least three times. Asterisks (*) indicate a significant difference compared with control groups (*P < 0.05, **P < 0.01 and ***P < 0.001). NC, negative control. KD, knockdown. (N) CALCR mediated expression of inflammatory factors in vitro. Experiments were repeated at least three times. Asterisks (*) indicate a significant difference compared with control groups (*P < 0.05, **P < 0.01 and ***P < 0.001).