Fig. 4

The up-regulation of HOXD3 expression predicts poor outcome in CRC patients and promotes CRC progression. a-b Expression analysis of HOXD-AS1 in normal colorectal mucosa and CRC tissues by a IHC and b the statistical analysis for HOXD3 expression. c Graphical illustration of statistical HOXD3 distribution in CRC patients. d-f Kaplan–Meier analysis of overall survival in all patients with CRC according to HOXD3 expression in clinical samples (overall survival, n = 164, log-rank test) and TCGA database (overall survival and disease-free survival, n = 360, log-rank test). g Infecting M5 and SW620 cells with a lentivirus vector harbouring shRNA-HOXD3 to knocked down the endogenous expression of HOXD3 in cells. HOXD3 levels in cells were detected by real-time PCR. h HOXD3 levels in cells were detected by Western blot. i CCK-8 assays were performed to determine the proliferation of HOXD3- depleted CRC cells. j Colony-forming assays were performed to determine the effects of HOXD3 depletion on the growth of CRC cells. The diameter > 50 cells was scored. k Cell cycle progression was analyzed by flow cytometry. l The migration potencies of CRC cells with the indicated treatments were detected by using wound healing assay. m Invasion assays were used to determine the effects of HOXD3 depletion on the invasion ability of CRC cells. For b, g, i, j, k, l and m, data are presented as means ± SD in three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001