Network analysis relevant to GRB2-mediated HuCCT1 migration.

(A) Venn diagrams showing the significantly different mRNA expression levels of Pre-miR-376c and siGRB2-2 transfectants of HuCCT1 relative to appropriate controls. Expression profiles of mRNAs affected by Pre-miR-376c and siGRB2-2 in EGF-treated HuCCT1 cells were conducted by microarray analysis. The numbers of genes regulated by Pre-miR-376c and siGRB2-2 are indicated. (B) The network of the identified molecules regulated by both Pre-miR-376c and siGRB2-2 in this study were connected with EGF, EGFR and GRB2 by IPA analysis. Numbers below the upregulation (red) and downregulation (green) symbols represent the fold changes by Pre-miR-376c treatment; numbers in parentheses represent the fold changes by siGRB2-2 treatment. Solid and dotted lines indicate direct and indirect gene relationships, respectively. (C and D) Real-time PCR analysis of IL1B (C) and MMP9 (D) expression levels in EGF-treated HuCCT1 cells after transfection with Pre-miR-376c and siGRB2-2. Pre-miR-negative control and nonspecific non-silencing siRNA were used as negative controls (NC). For quantitative comparisons, expression levels were normalized to GAPDH. The expression levels in negative controls were set to 1.0. The significance of differences between means was determined by Student's t-test. * p<0.05.