Fig. 2
DDX27 promoted CRC cells proliferation and inhibited apoptosis. a The mRNA expression of DDX27 in seven CRC cell lines (DLD-1, LoVo, HT29, HCT-116, SW1116, SW480, SW620), a normal human colon epithelial cell line (NCM460) and human normal colon tissue was shown. b Western blot analysis of isolated nucleus and cytoplasm from four different CRC cell lines indicated that DDX27 was mainly localized in nucleus. Lamin A/C and α-tublin were used as nuclear marker and cytoplasmic marker, respectively. c Immunofluorescence staining for DDX27 (red), F-Actin (green) and DAPI (blue) was shown using HCT116 and SW480 cells. d HCT116 and SW480 were stably transduced with lentiviral vectors carrying DDX27 cDNA or DDX27-specific small hairpin RNAs (shDDX27). Accordingly, control groups were transduced with lentiviral carrying empty vector (EV) or negative control shRNA (shCTL). Overexpression or silencing of DDX27 was confirmed by RT-PCR (Top panel) and western blot analysis (Bottom panel). e Ectopic expression of DDX27 significantly promoted cell viability by MTT assay (top panel) and increased colony numbers (bottom panel) as compared to control groups in HCT116 and SW480. f Knockdown of DDX27 significantly reduced cell viability by MTT assay (top panel) and decreased colony numbers (bottom panel) as compared to controls in HCT116 and SW480. g and h HCT116 and SW480 cells with altered expression of DDX27 (overexpression or silencing) were treated with STS (1 and 1.5 uM, respectively). Apoptosis was analyzed by flow cytometry. The apoptotic index in the right panel was defined as the percentage of apoptotic cells. After STS treatment for 3 h, expression of the cleaved form and total level of caspase-8, caspase-3 and PARP was detected by western blot analysis. (*P < 0.05; **P < 0.01; ***P < 0.001)
