Figure 4
AEG-1 knockdown down-regulated MMP1 expression in HNSCC cells in vitro and in vivo . (A) microarray analysis of gene expression after AEG-1 knockdown in SAS cells. Genes with an absolute fold change greater than 2.5 are shown. SNORD3B-1, Homo sapiens small nucleolar RNA, C/D box 3B-1; EMILIN2, elastin microfibril interfacer 2; FABP4, fatty acid binding protein 4; MMP1, matrix metallopeptidase 1; ANO1, anoctamin 1; RELN, reelin; FPR2, formyl peptide receptor 2; UHMK1, U2AF homology motif kinase 1; RMND5A, required for meiotic nuclear division 5 homolog A; CAPZA1; capping protein (actin filament) muscle Z-line, alpha 1. *, p < 0.05; **, p < 0.01; ***, p < 0.001. (B) RT-QPCR confirmation of gene expression profiles after knockdown of AEG-1 in SAS and FaDu cells. All experiments were performed in triplicate (n = 3) and data were normalized to GAPDH. (C) secreted MMP1 protein in cell-conditioned culture media. Each cell lines were seeded in equal numbers (1 × 106 cells) and were cultured in starving condition (1% fetal bovine serum, FBS) for 24 hr and the media were harvested. (D) immunohistochemical staining of MMP1 in murine subcutaneous xenograft tumors (upper row) and metastatic lesions from in vivo lung metastasis assays (lower row). Scale bar: upper row, 100 μm; lower row, 25 μm.