Experimental Studies Butorphanol on the Biological Activity and Chemotherapy Drug Resistance of Osteosarcoma Cells by Regulating the FOXO3-FOXM1 Signal Axis
10.3969/j.issn.1671-7414.2024.06.006
- VernacularTitle:布托啡诺调节FOXO3-FOXM1信号轴对骨肉瘤细胞生物活性和化疗药物耐药性的实验研究
- Author:
Tingting AO
1
;
Zhenzhen JIANG
;
Xi ZHANG
Author Information
1. 十堰市人民医院麻醉科,湖北 十堰 442000
- Keywords:
butorphanol;
forkhead box protein O3-forkhead box protein M1 signaling pathway;
osteosarcoma;
cisplatin
- From:
Journal of Modern Laboratory Medicine
2024;39(6):37-42,66
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the impacts of butorphanol(BUT)on the biological activity and chemotherapy drug resistance of osteosarcoma cells by regulating the forkhead box protein O3(FOXO3)-forkhead box protein M1(FOXM1)signal axis.Methods CDDP resistant MG-63 cells(MG-63/CDDP)treated with 2.0 μmol/L cisplatin(CDDP)were separated into control group(MG-63/CDDP cells were treated with 0.05％DMSO medium),BUT group(MG-63/CDDP cells were treated with 40 μg/mL BUT),JY-2 group(MG-63/CDDP cells were treated with 100 μmol/L FOXO3-FOXM1 inhibitor JY-2),and BUT+JY-2 group(MG-63/CDDP cells were treated with 40 μg/ml BUT and 100 μmol/L JY-2).CCK8 method was applied to detect MG-63/CDDP cell activity.Flow cytometry was used to detect apoptosis in MG-63/CDDP cells.The Transwell method was applied to detect the migration and invasion of MG-63/CDDP cells;Western blot was applied to detect the expression of autophagy proteins and proteins related to the FOXO3-FOXM1 signaling pathway.Results Compared with MG-63 cells,the IC50(20.56±2.52μmol/L vs(0.97±0.10μmol/L)of MG-63/CDDP cells was increased,and the differences was statistically significant(q=19.017,P<0.05),and the optimal concentration of 1 μmol/L CDDP was selected for subsequent experiments.Compared with the control group,the A value(0.43±0.05 vs 0.68±0.06),numbers of cell migration(63.63±7.58 vs 114.56±10.57)and invasion(43.38±4.58 vs 79.56±8.48),and the levels of autophagy-related protein Beclin1(0.31±0.05 vs 0.62±0.07)and microtubule-associated protein light chain 3(LC3)-Ⅱ/Ⅰ proteins(0.51±0.08 vs 0.98±0.11)in the BUT group were reduced(q=6.763～9.591,all P<0.05),the apoptosis rate(28.57％±3.14％vs 8.67％±1.46％),the levels of FOXO3(0.72±0.08 vs 0.33±0.04)and FOXM1(1.22±0.15 vs 0.70±0.08)proteins were increased(q=14.077,10.681,7.493,all P<0.05),however,in the JY-2 group,the A value(0.99±0.13 vs 0.68±0.06),numbers of cell migration(147.59±15.37 vs 114.56±10.57)and invasion(111.83±12.58 vs 79.56±8.48),and the levels of Beclin1(0.94±0.11 vs 0.62±0.07)and LC3-Ⅱ/Ⅰ(1.27±0.13 vs 0.98±0.11)proteins in the JY-2 group were increased(q=4.171～6.012,all P<0.05),the apoptosis rate(4.56％±0.86％vs 8.67％±1.46％),the levels of FOXO3(0.17±0.01 vs 0.33±0.04)and FOXM1(0.46±0.03 vs 0.70±0.08)proteins were reduced(q=5.941,9.505,6.881,all P<0.05),and the differences were statistically significant,respectively.JY-2 reversed the beneficial effects of BUT on MG-63/CDDP cell activity and chemotherapy resistance.Conclusion BUT may regulate the cell activity and CDDP resistance of osteosarcoma cells by activating the FOXO3-FOXM1 signaling pathway.
