MicroRNA-15a - cell division cycle 42 signaling pathway in pathogenesis of pediatric inflammatory bowel disease

Figure 4 MicroRNA-15a negatively regulates epithelial junctions through cell division cycle 42 in Caco-2 cells. Caco-2 cells were stimulated by Tumor necrosis factor-α (TNF-α) (35 ng/mL) [wild type (WT) + TNF-α group] or medium with no TNF-α (WT). Caco-2 cells were transfected with miR-15a mimic lentivirus or its negative controls (miR-NC group). Caco-2 cells were transfected with microRNA-15a (miR-15a) inhibitor lentivirus, and after passage, were stimulated by TNF-α (35 ng/mL) (miR-15a inhibitor + TNF-α). Caco-2 cells were transfected with Cdc42 lentivirus (Lv-Cdc42) or its negative control (Lv-Cdc42-NC), and after passage, stimulated by TNF-α (35 ng/mL) (Lv-Cdc42 +TNF-α or Lv-Cdc42nc + TNF-α group). A: Quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to evaluate the relative mRNA expression of E-cadherin (E-Cad) in every group; B: Expression of epithelial marker E-cad was examined by immunofluorescence in every group. Cells were stained with E-cad antibodies and counter stained with DAPI; C: qRT-PCR was used to evaluate the relative mRNA expression of ZO-1 in every group; D: Expression of epithelial marker ZO-1 was examined by immunofluorescence in every group. Cells were stained with E-cad antibodies and counterstained with DAPI (^aP < 0.05, ^bP < 0.01 vs WT, ^cP < 0.05, ^dP < 0.01 vs miR-15a mimic, ^eP < 0.05 vs Lv-Cdc42-NC +TNF-α) (magnification 40 ×, bar = 25 μm). Cdc42: Cell division cycle 42; WT: Wildtype; miR: MicroRNA; NC: Negative control; TNF-α: Tumor necrosis factor-α; LV: Lentiviral; NC: Negative control; qRT-PCR: Quantitative reverse transcription polymerase chain reaction; ZO-1: Zona occludens-1.