Fig. 3: CircZNF566 promotes HCC progression by serving as an miRNA sponge of miR-4738-3p. | Cell Death & Disease

Fig. 3: CircZNF566 promotes HCC progression by serving as an miRNA sponge of miR-4738-3p.

From: Circular RNA circZNF566 promotes hepatocellular carcinoma progression by sponging miR-4738-3p and regulating TDO2 expression

Fig. 3

a, b The relative abundances of 10 miRNA candidates in the Huh7 and LM3 lysates with circZNF566 or obligo probe were examined by qRT-PCR. c Luciferase activities of luc-circZNF566 in HCC cells transfected with five selected miRNA mimics or NC mimic were determined by a luciferase reporter assay. d Wound healing and transwell migration and invasion assays were performed to analyze the mobility, migration and invasion, respectively, of Huh7 cells transfected with five selected miRNA mimics or NC mimic. e The obviously negative correlation between the levels of circZNF566 and miR-4738-3p in 57 pairs of fresh frozen HCC tissues and matched normal liver tissues was analyzed by Pearson correlation analysis. f Relative miR-4738-3p expression in 57 pairs of fresh frozen HCC tissues and matched normal liver tissues. g The relative levels of miR-4738-3p in Huh7 and LM3 cell lines transfected with si-circZNF566, circZNF566 or NC were detected by qRT-PCR. h Schematic illustration of the sequence of wild-type TDO2 3’-UTR (WT) and TDO2 3’-UTR with mutations at the miR-4738-3p binding sites (mutant). i The effects of miR-4738-3p mimics, inhibitor and NC on luciferase activity were detected in HCC cells transfected with luciferase reporter plasmids expressing the WT 3’-UTR, mutant 3’-UTR or NC. j RIP assay for circZNF566 was performed with an anti-AGO2 antibody in HCC cells transfected with mimics or NC, and the expression of circZNF566 and miR-4738-3p was detected by qRT-PCR. k The effects of circZNF566 and miR-4738-3p on the protein expression of Bcl-2, Ki67, Caspase2, N-cadherin, E-cadherin, and Vimentin were detected by WB. l The effects of circZNF566 and miR-4738-3p on Huh7 cells migration, invasion and proliferation were evaluated by the transwell migration and invasion assays, the colony formation assay and the CCK-8 assay. All data are from three independent experiments and are presented as the means ± SEM or representative of three independent experiments with similar results (*p < 0.05, **p < 0.01, ***p < 0.001).

Back to article page