Hsa_circRNA_102610 upregulation in Crohn’s disease promotes transforming growth factor-β1-induced epithelial-mesenchymal transition via sponging of hsa-miR-130a-3p

Figure 7 Overexpression of hsa_circRNA_102610 promoted mothers against decapentaplegic homolog 4 and CyclinD1 protein expression, and transforming growth factor-β1-induced epithelial-mesenchymal transition in human intestinal epithelial cells and normal-derived colon mucosa cell line 460 (NCM460) cells. A: Western-blot detection of mothers against decapentaplegic homolog 4 and CyclinD1 expression following hsa_circRNA_102610 downregulation or overexpression in human intestinal epithelial and NCM460 cells. Downregulation of hsa_circRNA_102610 was conducted by siRNA (Si-1, Si-2), and a hsa-miR-130a-3p inhibitor was included to reverse the effects of the siRNAs (Si1-IN, Si2-IN). Si-NC was the negative control for siRNA interference. Hsa_circRNA_102610 was overexpressed after pLC5-ciR plasmid transfection, and hsa-miR-130a-3p mimics were included to reverse the effect of hsa_circRNA_102610 overexpression (CirOE-Mir). Cirtrl was the overexpression negative control pLC5-ciR null vector transfection; B: human intestinal epithelial cell and normal-derived colon mucosa cell line 460 (NCM460) cell morphology following transforming growth factor-β1 (TGF-β1) treatment. The cell-cell junctions were reduced in TGF-β1 treated cells, and hsa_circRNA_102610 overexpression promoted this effect. Conversely, hsa-miR-130a-3p mimics protected cells from morphological changes, and hsa_circRNA_102610 overexpression reversed this effect; C: Western blot detection of mothers against decapentaplegic homolog 4, N-Cadherin, E-Cadherin, and Vimentin in human intestinal epithelial cells and NCM460 cells following TGF-β1 treatment. CIR OE: hsa_circRNA_102610 overexpression by pLC5-ciR plasmid transfection. MIR mimic: Transfection with hsa-miR-130a-3p mimics; SMAD4: Mothers against decapentaplegic homolog 4; Cir OE: Hsa_circRNA_102610 was overexpressed after pLC5-ciR plasmid transfection; CirOE-Mir: Hsa-miR-130a-3p mimics were included to reverse the effect of hsa_circRNA_102610 overexpression; TGF-β1: Transforming growth factor-β1. Bar: × 100 μm.