Functional role of long non-coding RNA CASC19/miR-140-5p/CEMIP axis in colorectal cancer progression in vitro

Figure 4 Cancer susceptibility 19 acts as a competing endogenous RNA by sponging miR-140-5p to regulate cell migration inducing hyaluronidase 1 expression. A: Differential mRNA expression of cell migration inducing hyaluronidase 1 (CEMIP) in colorectal cancer (CRC) tissues and adjacent healthy colonic mucosa tissues was determined by quantitative real-time PCR (qRT-PCR). B: Differential protein expression of CEMIP in CRC tissues and adjacent healthy colonic mucosa tissues was analyzed using Western blot and immunochemistry. C: Differential mRNA expression of CEMIP in CRC cell lines and FHC cells was determined by qRT-PCR. D: Cancer susceptibility 19 (CASC19) and CEMIP correlated positively in CRC tissues, based on Pearson’s correlation curve. E: Protein expression of CEMIP in SW480 and HCT-116 cells was determined by Western blot after transfection with CASC19-p or siCASC19. The grayscale value of protein was measured with Image J. F: Predicted binding sites of miR-140-5p in the CASC19 sequence. G: Differential expression of miR-140-5p in CRC tissues and adjacent healthy colonic mucosa tissues, CRC cell lines, and FHC cells was determined by qRT-PCR. H: CASC19 and miR-140-5p correlated negatively in CRC tissues, as determined by Pearson’s correlation curve. I: RNA-IP was performed in SW480 and HCT-116 cells and CASC19 expression was determined by qRT-PCR. J: Luciferase reporter assay verified miR-140-5p to be a direct target of CASC19. K: Predicted binding sites of miR-140-5p in the CEMIP mRNA sequence. L: CEMIP and miR-140-5p correlated negatively in CRC tissues, based on Pearson’s correlation curve. M: Luciferase reporter assay verified CEMIP to be a direct target of miR-140-5p. N: CEMIP expression in SW480 and HCT-116 cells transfected with miR-140-5p mimic or siCASC19 was determined by Western blot and qRT-PCR. O: The expression of CEMIP in SW480 and HCT-116 cells transfected with miR-140-5p inhibitor or CASC19-p was determined by Western blot and qRT-PCR. Data are reported as the mean ± SD. ^aP < 0.05. CASC19: Cancer susceptibility 19; CRC: Colorectal cancer; CEMIP: Cell migration inducing hyaluronidase 1.