Fig. 5

MiR-218-5p inhibited pancreatic cancer cell progression by directly regulating LASP1. a, The expression levels of miR-218-5p were detected by qRT-PCR in BxPC-3 and SW1990 cells transfected with the miR-218-5p mimic (miR-218-5p) or mimic control (con). U6 was used as a loading control. b, The expression levels of LASP1 protein in BxPC-3 and SW1990 cells transfected with the miR-218-5p mimic (miR-218-5p) or mimic control (con) were analyzed by Western blot. c, Representation of the seed pairing between miR-218-5p and LASP1 3’UTR. Three mutations (mut1-LASP1, mut2-LASP1 and mut3-LASP1) were generated for luciferase assay in the sequence complementary to the miR-218-5p target binding region. d, BxPC-3 and SW1990 cells were co-transfected with either miR-218-5p mimic (miR-218-5p) or mimic control (con) and the wild-type or mutant LASP1 3’UTR luciferase reporter vector for 48 h.The relative luciferase activity was determined by the dual-luciferase assay. Renilla luciferase activity was used as a loading control. e, Efficiency of LASP1 re-expression was determined by Western blot. f, CCK-8 analysis revealed that LASP1 re-expression partly reversed the growth repression of miR-218-5p on pancreatic cancer cells. g, The restoration of LASP1 expression reversed the suppressive effects of miR-218-5p in colony formation. h and I, Ectopic expression of LASP1 reversed the suppressive effects of miR-218-5p in migration and invasion. *P < 0.05, **P < 0.01