Potential cyclooxygenase-2-independent mechanisms of the anti-proliferation and anti-fibrotic effects of celecoxib on the human intrahepatic biliary epithelial cell line HIBEpiC assessed by bulk-RNA sequencing

This experiment utilized next-generation sequencing (NGS) to identify potential cyclooxygenase-2-independent mechanisms of the anti-proliferation and anti-fibrotic effects of celecoxib on the human intrahepatic biliary epithelial cell line HIBEpiC. HIBEpiC cells were cultured in vitro. Treatments were vehicle, 20 ng/mL of TGF-β (T20), combination of 10μM of celecoxib plus 20 ng/mL of TGF-β (C10+T20), or combination of 10μM of 2,5-dimethyl-celecoxib plus 20 ng/mL of TGF-β (DC10+T20) before NGS. The gene expression were analyzed. Overall, the results suggested that celecoxib had a COX-2-independent inhibitory effect on the proliferation and profibrotic behavior of TGF-β-stimulated CXCL12+ HIBEpiC cells, probably because of the regulation of the cell cycle and several other signaling pathways. HIBEpiC cells were cultured in vitro. Treatments were vehicle, 20 ng/mL of TGF-β (T20), combination of 10μM of celecoxib plus 20 ng/mL of TGF-β (C10+T20), or combination of 10μM of 2,5-dimethyl-celecoxib plus 20 ng/mL of TGF-β (DC10+T20). Vehicle: HIBEpiC cells were cultured in RPMI 1640 medium with 0.5% Bovine Serum Albumin (BSA) and 0.1% DMSO for 28 hours. T20: HIBEpiC cells were cultured in RPMI 1640 medium with 0.5% BSA and 0.1% DMSO for 4 hours, and then the TGF-β were added to the final concentration of 20 ng/mL for further 24 hours incubation. C10+T20: HIBEpiC cells were cultured in RPMI 1640 medium with 0.5% BSA and 10μM of celecoxib (final concentration of DMSO: 0.1%) for 4 hours, and then the TGF-β were added to the final concentration of 20 ng/mL for further 24 hours incubation. DC10+T20: HIBEpiC cells were cultured in RPMI 1640 medium with 0.5% BSA and 10μM of 2,5-dimethyl-celecoxib (final concentration of DMSO: 0.1%) for 4 hours, and then the TGF-β were added to the final concentration of 20 ng/mL for further 24 hours incubation. The cell experiments were performed in triplicate. Then, the cells were harvested for NGS.