Fig. 5 | Oncogene

Fig. 5

From: DEAD-box helicase 27 promotes colorectal cancer growth and metastasis and predicts poor survival in CRC patients

Fig. 5The alternative text for this image may have been generated using AI.

DDX27 enhanced and prolonged NF-кB signaling. a In left panel, different cancer pathway reporters consisting of specific pathway-focused transcription factor-responsive firefly luciferase construct were separately transfected to HCT116 cells along with renilla luciferase reporter as internal control. Right panel showed knockdown of DDX27 significantly inhibited NF-кB luciferase reporter activity in both HCT116 and SW480 cells. Reporter activity was determined as the ratio of firefly to Renilla luciferase activity. b Caffeic acid phenethyl ester (CAPE) or 4-methyl-1-N- (3-phenylpropyl) benzene-1,2-diamine (JSH-23) was added to cells at indicate concentration (For HCT116, 2.5 mg/L CAPE or 10 uM JSH-23 was used; For SW480, 10 mg/L CAPE or 20 uM JSH-23 was used). Control groups were treated with an equivalent dilution of DMSO. Cell viability was assessed by MTT assays. In both experimental groups (CAPE or JSH-23) and control groups (DMSO), the difference between growth rates of cells over-expressing DDX27 and cells carrying empty vector (EV) was determined by ANOVA with repeated-measures analysis of variances. c HCT116 cells stably overexpressing DDX27 and control cells were treated with 50 ng/ml TNF-α for 3 h. NFkB Signaling Pathway Plus PCR Array (Qiagen) which profiles the expression of 84 key genes related to NF-кB-mediated signal transduction was used to analyze the effect of DDX27 on the NF-кB signaling. The cutoff fold-change was set to 1.2. d and e SW480 or HCT116 cells were treated with 50 ng/ml TNF-α for indicated time. The mRNA expression levels of NF-кB target genes (BIRC3, CCL20, CXCL3, NFKBIA, TNF, and TNFAIP3) in SW480-EV and SW480-DDX27 (or HCT116-shCTL and HCT116-shDDX27) cells were measured by qPCR. Student’s t-test was performed between SW480-EV and SW480-DDX27 (or HCT116-shCTL and HCT116-shDDX27-2) cells at different time points. f SW480 or HCT116 cells stably overexpressing DDX27 and control cells were treated with 50 ng/ml TNF-α for indicated time. Isolated nucleus and cytoplasm were immunoblotted with indicated antibodies. Lamin A/C and α-tublin were used as nuclear marker and cytoplasmic marker, respectively. (*P < 0.05; **P < 0.01; ***P < 0.001)

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