Figure 5.

COL1A1 is a direct target of hsa-miR-193a-5p. (A) Venn diagram showing 75 genes that are putative hsa-miR-193a-5p targets computationally predicted by three algorithms (miRWalk, miRDB, and TargetScan). (B) These computed results were compared with the upregulated genes in TCGA (P < 0.05, logFC > 1). (C,D) A specific biotin-labeled miR-149-5p probe was used to capture COL1A1 successfully relative to the NC group. (E) A dual-luciferase reporter assay was performed to determine the direct binding between COL1A1 and hsa-miR-193a-5p based on their complementary sequences. (F) Increased expression of COL1A1 was observed in TCGA (expression matrix is normalized and log transformed). (G) The increased relative levels of COL1A1 were confirmed in the colorectal carcinoma tissues and corresponding adjacent tissues by qRT-PCR and normalized to GAPDH levels (N = 60, P < 0.001). (H) Relative mRNA levels of COL1A1 in CRC cells were determined by qRT-PCR in LOVO and HT29 cell lines transfected with the indicated plasmids. (I) COL1A1 protein levels in CRC cells were determined by qRT-PCR in LOVO and HT29 cell lines transfected with the indicated plasmids. Data are presented as the means ± standard deviation (**P < 0.01).