Effects of Remimazolam on the Proliferation, Apoptosis, and Epithelial-Mesenchymal Transition of Bladder Cancer Cells by Regulating the HMGB1-RAGE Pathway
WU Xiaoxu1, LUO Shujun1, WANG Ran1, WU Anshi2*
This study aims to investigate the mechanism by which remimazolam regulates the HMGB1-RAGE pathway and its subsequent effects on the proliferation, apoptosis, and EMT (epithelial-mes enchymal transition) of bladder cancer cells. The immortalized bladder cell line SV-HUC-1 and the bladder cancer cell line BIU-87 were cultured, and cell viability following remimazolam treatment was assessed us ing the CCK-8 assay. BIU-87 cells were sequentially divided into the following groups: control group, low dose remimazolam group, medium-dose remimazolam group, high-dose remimazolam group, and high-dose remimazolam+rHMGB1 (recombinant human HMGB1) group. Cell proliferation capacity was evaluated by colony formation assay; cell apoptosis was detected by AO/EB double staining and Annexin V-FITC/PI double staining; EMT was detected by immunofluorescence; the levels of cell behavior-associated proteins and HMGB1-RAGE pathway-associated proteins were detected by Western blot. Compared with the 0 µg/mL remimazolam group, the viability of BIU-87 cells was significantly decreased when the concentration was in the range of 0.5 µg/mL to 4 µg/mL (P<0.05). Compared with the control group, the number of colonies formed, Ki-67 positive cell rate, Fibronectin positive cell rate, p-p65/p65 ratio, and the protein levels of c-myc, CyclinD1, XIAP, Bcl-2, HMGB1, and RAGE were decreased in the low-, medium-, and high-dose remima zolam groups, while the cell apoptosis rates detected by AO/EB double staining and Annexin V-FITC/PI flow cytometry, and the positive cell rates of E-cadherin and Claudins were increased. The inhibitory effect of the high-dose remimazolam group on malignant behaviors of BIU-87 cells was significantly stronger than those of the medium- and low-dose groups (P<0.05). Compared with high-dose remimazolam treatment alone, high-dose remimazolam combined with rHMGB1 partially reversed the inhibitory effect of remimazolam on malignant behaviors of bladder cancer cells (P<0.05). Remimazolam regulates the proliferation, apoptosis, and EMT process of bladder cancer cells by inhibiting the activation of the HMGB1-RAGE pathway.
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