Research Paper Volume 13, Issue 7 pp 9838—9858

Figure 4. miR-590-3p binds to SNHG4 and the CDK1 3'-UTR. (A) The online tools mirDIP and R programming language were used to predict miRNAs that might simultaneously target SNHG4 and the CDK1 3'-UTR, and miR-590-3p was identified. (B) The expression of miR-590-3p was determined in 12 CRC and normal noncancerous tissues by real-time PCR. (C) The expression of miR-590-3p was determined in one normal fetal colon cell line (FHC) and five CRC cell lines (HCT8, LoVo, HCT116, SW620, and HT29) by real-time PCR. (D) miR-590-3p was overexpressed or inhibited in HCT116 and SW620 cells by transfection with miR-590-3p or anti-miR-590-3p, and the effects were confirmed by real-time PCR. (E) HCT116 and SW620 cells were transfected with miR-590-3p or anti-miR-590-3p, and the mRNA levels of SNHG4 were examined by real-time PCR. (F) HCT116 and SW620 cells were transfected with miR-590-3p or anti-miR-590-3p, and the protein levels of CDK1 were examined by immunoblotting. (G–H) Luciferase reporter assays were performed by constructing luciferase reporter vectors, as described in the Materials and methods section, to validate the predicted binding of miR-590-3p to SNHG4 and the CDK1 3'-UTR. ^*P < 0.05, ^**P < 0.01, ^##P < 0.01.