依托咪酯通过miR-125b-5p介导JAK/STAT途径抑制膀胱癌细胞恶性生物学行为的机制研究OA

Objective To investigate the effect of etomidate on bladder cancer cells through microRNA-125b-5p(miR-125b-5p)and its mechanism.Methods BIU-87 cells were divided into control group,experimental group(cultured with 450 mg·L-1 etomidate for 24 h),miR-NC group(transfected with miR-NC plasmid,cultured with 450 mg·L-1 etomidate for 24 h),miR-125b-5p inhibitor group(transfected with miR-125b-5p inhibitor plasmid,cultured with 450 mg·L-1 etomidate for 24 h).Quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression level of miR-125b-5p in each group of cells,cell counting kit-8(CCK-8)was utilized to evaluate the proliferative capacity of cells in each group,cell scratch assay was used to determine the migratory ability of cells in each group,Transwell assay was adopted to assess the invasive potential of cells in each group,immunofluorescence was used to detect the level of epithelial-mesenchymal transition(EMT)-related proteins in each group of cells and Western blot was used to detect the level of Janus kinase 2(JAK2)/signal transducer and activator of transcription(STAT3)pathway-related proteins in each group of cells.Results The relative expression levels of miR-125b-5p in the control group,experimental group,miR-NC group and miR-125b-5p inhibitor group were 1.00±0.14,2.72±0.52,2.65±0.48 and 1.58±0.21,respectively;the 72 h cell proliferation rate were(100.00±12.87)％,(61.87±7.92)％,(64.58±10.92)％and(89.77±9.85)％,respectively;the migration ability were(68.21±7.25)％,(23.85±3.92)％,(22.67±3.58)％and(45.13±6.85)％,respectively;the number of invasions were(132.56±15.88),(61.58±11.85),(60.89±10.92)and(92.37±13.85)cells,respectively;the relative expression levels of epithelial-cadherin(E-cadherin)protein were 1.00±0.15,1.45±0.22,1.32±0.12 and 1.14±0.16,respectively;the relative expression levels of neural-cadherin(N-cadherin)protein were 1.00±0.13,0.55±0.06,0.51±0.06 and 0.59±0.05,respectively;the relative expression levels of vimentin(Vim)protein were 1.00±0.16,0.48±0.08,0.51±0.05 and 0.61±0.06,respectively;the relative expression levels of phosphorylated(p)-JAK2/JAK2 protein were 1.00±0.15,0.57±0.08,0.54±0.07 and 0.76±0.09,respectively;the relative expression levels of p-STAT3/STAT3 protein were 1.00±0.13,0.67±0.08,0.64±0.07 and 0.86±0.09,respectively.Compared the experimental group with the control group,and compared the miR-125b-5p inhibitor group with the miR-NC group,the differences of above indicators were all statistically significant(P＜0.05,P＜0.01,P＜0.001).Conclusion Etomidate inhibits the proliferation,migration,invasion,and EMT of BIU-87 cells by up-regulating miR-125b-5p,which may be achieved by inhibiting the JAK2/STAT3 pathway.