Role of microRNA-30c Targeting ADAM19 in Colorectal Cancer
Fig 4
ADAM19 was a direct target of miR-30c.
(A) The predicted targeting site with miR-30c of ADAM19 3’-UTR; MiR-30c targeting sequences of ADAM19 3’-UTR are evolutionarily conserved through eight species (Hsa: human; Ptr: chimpanzee; Mml: rhesus; Oga: bushbabay; Tbe: treeshrew; Mmu: mouse; Rno: rat; and Ocu: rabbit.). The targeting sites are highlighted in red. (B-C) qRT-PCR and western blot analysis was applied to detect mRNA and protein expression of ADAM19 in HCT116 cells after transfection of miR-30c mimic or NC mimic. (D) ADAM19 mRNA and miR-30c expression level was determined in five colon cancer cell lines by qRT-PCR. (E) ADAM19 mRNA and miR-30c levels were inversely correlated in colon cancer tissues as determined by qRT-PCR. β-actin and U6 were used as the endogenous controls, respectively. (F) HEK293T cells were co-transfected with wild-type or mutant reporters and the miR-30c mimic or negative control (NC mimic). After 48h, Luciferase/Renilla activity was measured. (G) ADAM19 was upregulated in colon cancer tissues compared with normal tissues, as indicated by Western blotting.