Figure 5. LRRC1 was a direct target of miR-432-5p in CCA. (A) Targetscan, circinteractome and mirMap were used to identify potential mRNA target genes for miR-432-5p. (B) Relative expression of LRRC1 mRNA in CCA tissues and adjacent normal tissues. (C) qRT-PCR was used to detect the correlation between miR-432-5p and LRRC1 expression in CCA specimens. (D, E) The expression of LRRC1 mRNA and protein in CCLP-1 and QBC939. (F, G) The LRRC1 expression was detected in CCA cells with miR-432-5p up-regulation and down-regulation by qRT-PCR and western blot. (H) Diagrammatic sketch of the binding sites between miR-432-5p and LRRC1. (I) Luciferase reporter assay in CCA cells transfected with LRRC1 WT or LRRC1 MUT, along with miR-432-5p mimics or NC mimics. (J) RIP assay further verified the direct interaction between miR-432-5p and LRRC1. (K) Colony formation assay was used to detect the proliferation ability change after LRRC1 down-regulation and up-regulation. (L, M) The migration and invasion changes of tumor cells after LRRC1 down-regulation and up-regulation detected by transwell. *P< 0.05; **P< 0.01; ***P< 0.001.