Figure 5
In vitro functional analysis and expression of MMP14 and Snail in GC cells, and ectopic expression of MMP14 or Snail restores the effects of miR-22 on cell migration and invasion in GC cells. (a, b, d and e) qRT-PCR assays show the mRNA expression of MMP14 and Snail in GC cells transfected with si_con, si_MMP14#1, si_MMP14#2, si_Snail#1, si_Snail#2, pcDNA3.1 vector, pcDNA3.1-MMP14 vector, or pcDNA3.1-Snail vector, respectively. Data are presented as mean±S.D. (n=3). **P<0.01. (c and f) Western blotting assays show the protein expression of MMP14 and Snail in GC cells transfected with si_con, si_MMP14#1, si_MMP14#2, si_Snail#1, si_Snail#2, pcDNA3.1 vector, pcDNA3.1-MMP14 vector, or pcDNA3.1-Snail vector. GAPDH served as an internal reference. (g and h) Inhibition of cell migration and invasion by knockdown of MMP14 and Snail. SGC-7901 and HGC-27 cells were transfected with si_con, si_MMP14#1, si_MMP14#2, si_Snail#1, or si_Snail#2, respectively. The assays were repeated three times. Data are presented as mean±S.D. (n=5). **P<0.01. (i and j) Promotion of cell migration and invasion by overexpression of MMP14 or Snail. SGC-7901 and HGC-27 cells were transfected with pcDNA3.1 vector, pcDNA3.1-MMP14 vector, or pcDNA3.1-Snail vector, respectively. The assays were repeated three times. Data are presented as mean±S.D. (n=5). *P<0.05, **P<0.01. (k) Western blot assays show the protein expression of MMP14 and MMP2 in GC cells after transfection with miR-NC, miR-22 mimics, or co-transfection with miR-22 mimics and pcDNA3.1-MMP14 vector. GAPDH served as an internal reference. (l) Western blot assays show the protein expression of Snail and E-cadherin in GC cells after transfection with miR-NC, miR-22 mimics, or co-transfection with miR-22 mimics and pcDNA3.1-Snail vector. GAPDH served as an internal reference. (m and n) MMP14 or Snail overexpression partially rescues miR-22-reduced cell migration and invasion. Data are presented as mean±S.D. (n=5). **P<0.05. The assays were repeated in duplicate
