microRNA-627-5p inhibits colorectal cancer cell proliferation, migration and invasion by targeting Wnt2

Figure 1 Evaluation of the tissue expression and diagnostic utility of microRNA-627 in the GSE41655 and GSE18392 datasets. A: Tissue expression of microRNA-627 (miR-627) in the healthy controls (HCs) and colorectal cancer (CRC) patients in the GSE41655 dataset; B: Tissue expression of miR-627 in the HCs and CRC patients in the GSE18392 dataset; C: Receiver operating characteristic analysis of miR-627 to distinguish CRC patients from HCs in the GSE41655 dataset; D: Receiver operating characteristic analysis of miR-627 to distinguish CRC patients from HCs in the GSE18392 dataset. HCs: Healthy controls; CRC: Colorectal cancer. ^bP < 0.01; ^cP < 0.001.

Figure 2 Inverse correlation between Wnt2 and microRNA-627-5p expression in colorectal neoplasm tissues. A: The mRNA expression levels of Wnt2 in healthy control tissues, advanced adenoma tissues and colorectal cancer (CRC) tissues; B: The mRNA expression levels of Wnt2 in CRC cell lines (SW480, HCT116, and RKO cells) and epithelial cell line cell line; C: The relationship between miR-627-5p and Wnt2 mRNA expression in colorectal neoplasm tissues. HCs: Healthy controls; CRC: Colorectal cancer; AA: Advanced adenoma; FHC: Colonic epithelial cell line. ^cP < 0.001.

Figure 3 The direct binding relationship between microRNA-627-5p and Wnt2. A: Schematic illustration of the predicted binding sites between microRNA-627-5p (miR-627-5p) and Wnt2 mRNA; B: Dual luciferase reporter assays in HEK-293T cells. Experimental group: NC mimics + pcDNA-Wnt2-WT, miR-627-5p mimics + pcDNA-Wnt2-WT, NC mimics + pcDNA-Wnt2-MUT, miR-627-5p mimics + pcDNA-Wnt2-MUT; C: The transfection efficiency of miR-627-5p mimics in SW480 and HCT116 cells; D: The effects of miR-627-5p overexpression on the transcript expression levels of Wnt2 in SW480 and HCT116 cells; E: The effects of miR-627-5p overexpression on the protein expression levels of Wnt2 in SW480 and HCT116 cells. 3’ UTR: 3’-untranslated region; NS: Not significant; ^bP < 0.01; ^cP < 0.001.

Figure 4 Cellular behaviours induced by microRNA-627-5p mimics and Wnt2 overexpression plasmids in SW480 and HCT116 cells. Experimental group: NC mimics, microRNA-627-5p (miR-627-5p) mimics, miR-627-5p mimics + pcDNA-Wnt2. A and D: Scratch assay was used to detect the migration of colorectal cancer (CRC) cells in each group; B and E: Matrigel invasion assay was used to detect the invasive capability of CRC cells in each group; C and F: Flow cytometry analysis was used to evaluate the apoptosis of CRC cells in each group; G: Cholecystokinin octapeptide (CCK-8) assay was used to detect the viability of CRC cells in each group. NS: Not significant; CRC: Colorectal cancer. ^aP < 0.05; ^bP < 0.01; ^cP < 0.001.

Figure 5 Identification of the transfected efficiency of Wnt2 overexpression plasmids. A: The mRNA expressions of Wnt2 in SW480 and HCT116 cells after the transfection of Wnt2 overexpression plasmids; B: The protein expressions of Wnt2 in SW480 and HCT116 cells after the transfection of Wnt2 overexpression plasmids. ^bP < 0.01; ^cP < 0.001.

Figure 6 The mRNA expression alterations of downstream target genes in the Wnt/β-catenin signalling induced by miR-627-5p mimics and Wnt2 overexpression plasmids in SW480 and HCT116 cells. A: SW480 cells; B: HCT116 cells. Experimental group: NC mimics, miR-627-5p mimics, miR-627-5p mimics + pcDNA-Wnt2. NS: Not significant; ^bP < 0.01; ^cP < 0.001.

Figure 7 The protein expression alterations of downstream target genes in the Wnt/β-catenin signalling induced by microRNA-627-5p mimics and Wnt2 overexpression plasmids in SW480 and HCT116 cells. A: SW480 cells; B: HCT116 cells. Experimental group: NC mimics, miR-627-5p mimics, miR-627-5p mimics + pcDNA-Wnt2. NS: Not significant. ^aP < 0.05; ^bP < 0.01; ^cP < 0.001.