Figure 4.

Downregulation of UHRF1 induces upregulation of IL-6 promoting EMT. (a) qRT-PCR analysis for the mRNA expression levels of GRO, IL-6, IL-8 and MCP1 in shCont- and shUHRF1-HepG2 cells. (b) Effects of siRNAs targeting GRO, IL-6, IL-8 and MCP1 on the invasive property of shUHRF1-HepG2 cells. The cells were grown in the presence or absence of siRNAs for 48 h, and the invasion assay was performed using the Transwell chamber. (c) Effect of siRNA targeting IL-6 on the protein expression level of Snail in shUHRF1-HepG2 cells. The cells were grown in the presence or absence of siRNA targeting IL-6 for 48 h, and western blot analysis was performed. (d and e) Effects of a neutralizing antibody of IL-6 on the migratory and invasive properties of shUHRF1-HepG2 cells. The cells were grown in the presence or absence of a neutralizing antibody of IL-6 for 48 h, and the migration and invasion assay were performed using the Transwell chamber. (f) Effects of a neutralizing antibody of IL-6 on the protein expression level of Snail in shUHRF1-HepG2 cells. The cells were grown in the presence or absence of a neutralizing antibody of IL-6 for 48 h, and western blot analysis was performed. (g) Effect of siRNA targeting CXCR4 on the mRNA expression of IL-6 in shUHRF1-HepG2 cells. The cells were grown in the presence or absence of siRNA targeting CXCR4 for 48 h, and qRT-PCR analysis was performed. (h) Effect of siRNA targeting IL-6 on the mRNA expression of CXCR4 in shUHRF1-HepG2 cells. The cells were grown in the presence or absence of siRNA targeting IL-6 for 48 h, and qRT-PCR analysis was performed. (i) Effect of SDF1α (250 ng/ml) on the secretion of IL-6 from shUHRF1-HepG2 cells. The cells were grown in the presence or absence of SDF1α for 48 h, and ELISA was performed to detect the secretion of IL-6 using conditioned medium obtain from the cells treated with SDF1α. β-actin was used as a loading control. Results from three independent experiments are expressed as means ± SEMs. (*P < 0.05, **P < 0.01).