Fig. 2
MIR100HG drives cetuximab resistance and metastasis in CRC cells. a Mean CTX inhibition rate of CRC cells categorized by the gene expression–based consensus molecular subtyping (CMS). b Colony counts for 3D-cultured MIR100HGKOE4 cells in the presence or absence of CTX (3 μg/ml) after 18 d (n = 3 independent experiments performed in triplicate). c Representative images of Ki-67 (green) and cleaved Caspase-3 (red) staining in MIR100HGKOE4 CC-CR cells treated with CTX (10 μg/ml) for 24 h after culturing in 3D culture for 12 d, n = 5 independent experiments. Scale bar, 50 μm. d Colony counts for 3D-cultured CC-CR cells with indicated treatment (n = 3 independent experiments performed in triplicate). e Representative bioluminescence images of athymic nude mice (n = 7) injected subcutaneously with WT or MIR100HGKOE4 CC-CR cells that received CTX treatment (1 mg per mouse, intraperitoneal (i.p.) injection every 3 d). f, g Growth curve with image (f) and weight (g) of tumors in athymic nude mice (n = 7) injected with indicated cells. h Expression of MIR100HG in CRC patients with (M1) or without (M0) metastasis from TCGA database. i, j Extent of migration or invasion using Transwell migration and invasion assays of indicated cells when MIR100HG was overexpressed (i) and knocked down or knocked out (j). n = 3 independent biological replicates. k Representative bioluminescence images of indicated groups of mice (n = 10) at 8 weeks after tail vein injection (left) as well as radiance measurements (right). l Representative hematoxylin and eosin (H&E)-stained images of lung tissue sections from different groups (left). The number of lung metastatic foci was calculated (right) (n = 10). Scale bars, 200 μm. m, n Overall survival for mice in indicated groups. ***P < 0.001, **P < 0.01, *P < 0.05. Data represent mean ± s.d., n.s., not significant