AQP3 silence blocked PI3K/AKT

pathway in SGC7901 cells. AQP3 silence led to a significant decrease in phosphorylation of ser473 in AKT. * p<0.05 BLANK control SGC7901 cells NC cells treated with scrambled shRNA aqp3shRNA cells treated with aqp3shRNA AQP3 up-regulation activated PI3K/AKT pathway in SGC7901 cells We compared levels of phosphorylated and total AKT in SGC7901 cells with AQP3 over-expression by using Dorsomorphin Western blot. AQP3 over-expression led to a significant increase in phosphorylation of ser473 in AKT. (Figure 5) Figure 5 AQP3 regulated PI3K/AKT pathway in SGC7901 cells. AQP3 over-expression activated PI3K/AKT pathway in SGC7901 cells. AQP3 over-expression led to a significant increase in phosphorylation of ser473 in AKT. * p < 0.05 BLANK control SGC7901 cells NC cells treated with scrambled shRNA LV-AQP3 cells treated with lentiviral vector encoding AQP3 LY294002 down-regulated MMPs expression in SGC7901 cells SGC7901 cells were exposed to 20 μM LY294002 for 48 h (fresh media containing LY294002

was added every 24 h), and then were harvested to perform Western blot. We found a significant decrease in MT1-MMP, MMP-2, and MMP-9 expression. However, with Doramapimod the addition of LY294002, the expression of MMPs could not be obviously reversed in LV-AQP3 or aqp3shRNA groups. (Figure 6) Figure 6 LY294002 down-regulated MMPs expression and blocked the effect of LV-AQP3 and aqp3shRNA in SGC7901 cells. SGC7901 cells were exposed to LY294002 for 48h and then were harvested to perform Western blot analysis. We found a significant decrease in MT1-MMP, MMP-2, and MMP-9 expression. However, with the addition of LY294002, the expression of MMPs could not be obviously reversed in LV-AQP3 or aqp3shRNA groups. * p < 0.05 BLANK control SGC7901 cells NC cells treated with scrambled shRNA LY294002 cells treated with LY294002 LY294002+LV-AQP3 cells treated with LY294002 and LV-AQP3 all LY294002+aqp3shRNA cells treated with LY294002 and aqp3shRNA Discussion Recent

studies showed that the involvement of AQPs in angiogenesis and tumor cell migration and proliferation had potentially important clinical implication [10, 11]. We reported for the first time that AQP4 protein and mRNA expression levels in gastric cancer tissue were significantly lower than those in normal gastric tissue [12]. Then, we demonstrated that AQP3 played a critical role in gastric cancer cell migration and proliferation in previous study [13]. In this study, we found that AQP3 silence could down-regulate MMPs expression and AQP3 over-expression could up-regulate MMPs expression in SGC7901 cells. Many tumors exhibit GDC-0973 concentration elevated levels of MMPs, which may play an important role in cellular invasion and metastasis [14]. Among the human MMPs reported to date, MT1-MMP, MMP-2 and MMP-9 are the major enzymes involved in degrading types I and IV collagen and the extracellular matrix(ECM) [15].