Fig. 6

circPARD3 functions as a sponge of miR-145-5p in LSCC cells. a RIP assays were performed using an antibody against AGO2 with extracts from FD-LSC-1 and Tu 177 cells. Enrichment of circPARD3 in RNA samples after RIP assays was determined by qPCR analysis. b Venn diagram shows the intersection of predicted circPARD3 binding miRNAs and miRNAs downregulated in 107 LSCC tissues from the RNA sequencing data. c RNA pulldown assay was performed using Biotin-labeled circPARD3 probes in FD-LSC-1 cells, then the enrichment of the indicated miRNAs was detected by qPCR analysis. d Expression of miR-145-5p in LSCC cells overexpressing or silencing of circPARD3 was determined by qPCR analysis. e Luciferase reporter assay of FD-LSC-1 cells cotransfected with miR-145-5p mimics and luciferase reporter plasmid containing wild-type (WT) and miR-145-5p binding site mutated (Mut) circPARD3. N.S., no significant. f FD-LSC-1 and Tu 177 cells overexpressing circPARD3 (circPARD3-OE) were transfected with miR-145-5p mimics or NC mimics, expression of LC3B and p62 was detected by western blotting. g and h Cell proliferation of FD-LSC-1 and Tu 177 cells overexpressing circPARD3 and miR-145-5p was determined by CCK8 assays (g) and colony formation assays (h). The migration (i) and invasion (j) abilities of FD-LSC-1 and Tu 177 cells overexpressing circPARD3 and transfected with miR-145-5p mimics were evaluated by Transwell assays. Scale bar, 200 μm. k LSCC cells overexpressing circPARD3 and miR-145-5p mimics were treated with various concentrations of Cisplatin for 24 h. Cell viability was determined by CCK8 assays. Error bars represent SD of three independent experiments. *P < 0.05, **P < 0.01