MiR-301a transcriptionally activated by HIF-2α promotes hypoxia-induced epithelial-mesenchymal transition by targeting TP63 in pancreatic cancer

Figure 2 Upregulation of miR-301a promotes the epithelial-mesenchymal transition of pancreatic cancer cells. A: MiR-301a was upregulated in CFPAC-1 and BxPC-3 cells; B: MiR-301a upregulation promoted the transition of the CFPAC-1 and BxPC-3 cells from an epithelial morphology to a mesenchymal morphology; C and D: Western blot analysis was performed to detect the protein expression of hypoxia-inducible factor (HIF)-1α, HIF-2α, and the epithelial-mesenchymal transition markers; the expression of HIF-1α, HIF-2α, and the epithelial markers (E-cadherin and β-catenin) was upregulated in miR-301a-overexpressing cells, while the expression of the mesenchymal markers was downregulated; E and F: Immunofluorescence was performed to detect the protein expression of HIF-1α, HIF-2α, and the epithelial-mesenchymal transition markers in CFPAC-1 and BxPC-3 cells; the expression of HIF-1α, HIF-2α, and E-cadherin was upregulated in miR-301a-overexpressing cells, while the expression of the mesenchymal markers (Vimentin) was downregulated. HIF-1α protein accumulation in the nuclei of the tumor cells was more obvious in the miR-301a-overexpressing group than in the control group; G-J: Wound healing assays and Transwell assays were performed to measure CFPAC-1 and BxPC-3 migration; miR-301a overexpression improved CFPAC-1 and BxPC-3 migration under normoxia and hypoxia. ^bP < 0.01, ^cP < 0.001. HIF: Hypoxia-inducible factor.