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Fig. 3 | Cancer Cell International

Fig. 3

From: miR-199b-3p contributes to acquired resistance to cetuximab in colorectal cancer by targeting CRIM1 via Wnt/β-catenin signaling

Fig. 3

CRIM1 is a direct target of miR-199b-3p in acquired resistance to CTx in CRC. A Heatmap of the top 25 up-regulated genes and down-regulated genes between CTx treated tumors and CTx secondary resistant tumors based on GSE140973 dataset. B Venn diagram displaying only one down-regulated gene (CRIM1) of CTx secondary resistant tumors in the list of potential targets of miR-199b-3p. C Dual-luciferase reporter assay confirmed the interaction between miR-199b-3p and CRIM1 in SW480-CTxR and HCT116-CTxR (The putative binding sites were predicted using targetscan). D FISH assay displayed the intracellular distribution of miR-199b-3p in SW480-CTxR and HCT116-CTxR (Scale bars: 50 μm). E Anti-Ago2 RIP assay followed by qRT-PCR determined the association between miR-199b-3p and CRIM1 with Ago2 in SW480-CTxR and HCT116-CTxR. F Western blot examined the protein expression levels of CRIM1 in SW480-CTxR and HCT116-CTxR after the transfection of miR-199b-3p inhibitor or mimic. G The expression of CRIM1 in CRC tissues between responder and non-responder groups (n = 15 per group) using qRT-PCR (left); Pearson correlation analysis for miR-199b-3p and CRIM1 expression in CRC tissues from non-responders (n = 15) (right). H Representative images of CRIM1 expression in CRC tissues between responder (high expression) and non-responder (low expression) using IHC (Scale bars: 100 μm). I Western blot examined the protein expression levels of CRIM1 in CRC tissues between responder and non-responder groups. J The expression of CRIM1 between CRC-CTxR cell lines and the parental sensitive ones. K The correlation between the expression level of CRIM1 and IC50 for CTx in eight CRC cell lines (SW480-CTxR, HCT116-CTxR, HT29-CTxR, CaCO2-CTxR, SW480, HCT116, HT29, and CaCO2). ***P < 0.005 compared with responder or normal group

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