[1]奥婷婷,姜祯珍,张 熙.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[J].现代检验医学杂志,2024,39(06):37-42+66.[doi:10.3969/j.issn.1671-7414.2024.06.006]
　AO Tingting,JIANG Zhenzhen,ZHANG Xi.Experimental Studies Butorphanol on the Biological Activity and Chemotherapy Drug Resistance of Osteosarcoma Cells by Regulating the FOXO3-FOXM1 Signal Axis[J].Journal of Modern Laboratory Medicine,2024,39(06):37-42+66.[doi:10.3969/j.issn.1671-7414.2024.06.006]
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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()
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《现代检验医学杂志》[ISSN:/CN:]

卷:
第39卷
期数:
2024年06期
页码:
37-42+66
栏目:
论著
出版日期:
2024-11-15

文章信息/Info

Title:
Experimental Studies Butorphanol on the Biological Activity and Chemotherapy Drug Resistance of Osteosarcoma Cells by Regulating the FOXO3-FOXM1 Signal Axis
文章编号:
1671-7414（2024）06-037-07
作者:
奥婷婷; 姜祯珍; 张 熙
（十堰市人民医院麻醉科，湖北十堰442000）
Author(s):
AO Tingting; JIANG Zhenzhen; ZHANG Xi
(Department of Anesthesiology, Shiyan People’s Hospital, Hubei Shiyan 442000, China)
关键词:
布托啡诺; 叉头框蛋白O3- 叉头框蛋白M1 信号通路; 骨肉瘤; 顺铂
分类号:
R738.1；R730.43
DOI:
10.3969/j.issn.1671-7414.2024.06.006
文献标志码:
A
摘要:
目的 探究布托啡诺（BUT）调节叉头框蛋白O3（FOXO3）- 叉头框蛋白M1（FOXM1）信号轴对骨肉瘤细胞生物活性和化疗药物耐药性的影响。方法 将2.0μmol/L 顺铂（CDDP）处理的CDDP 耐药MG-63 细胞（MG-63/CDDP）分为对照组（MG-63/CDDP 细胞用含0.05g/dl DMSO 培养液处理）、BUT 组（40 μg/ml BUT 处理MG-63/CDDP 细胞）、JY-2 组（用100μmol/L FOXO3-FOXM1 抑制剂JY-2 处理MG-63/CDDP 细胞）和BUT+JY-2 组（用40μg/ml BUT 以及100μmol/L JY-2 处理MG-63/CDDP 细胞）。CCK8 法检测MG-63/CDDP 细胞活性；流式细胞术检测MG-63/CDDP 细胞凋亡情况；Transwell 法检测MG-63/CDDP 细胞迁移、侵袭情况；Western blot 检测自噬蛋白以及FOXO3-FOXM1 信号通路相关蛋白表达。结果 与MG-63 细胞相比，MG-63/CDDP 细胞IC50 增加（20.56±2.52μmol/L vs 0.97±0.10μmol/L），差异具有统计学意义（q=19.017，P<0.05），筛选出较适浓度1 μmol/L CDDP 用于后续实验。与对照组相比，BUT 组MG-63/CDDP 细胞A 值（0.43±0.05 vs 0.68±0.06），细胞迁移数量(63.63±7.58 个vs114.56±10.57 个) 以及侵袭数量(43.38±4.58 个vs 79.56±8.48 个)、自噬相关蛋白Beclin1(0.31±0.05 vs 0.62±0.07)和微管相关蛋白轻链3(LC3)-II/I 蛋白(0.51±0.08 vs 0.98±0.11) 水平均下降（q=6.763 ～ 9.591，均P<0.05），凋亡率（28.57%±3.14% vs 8.67%±1.46%），FOXO3（0.72±0.08 vs 0.33±0.04），FOXM1（1.22±0.15 vs 0.70±0.08）蛋白水平均上升（q=14.077，10.681，7.493， 均P<0.05）， 而JY-2 组MG-63/CDDP 细胞A 值（0.99±0.13 vs0.68±0.06），细胞迁移数量(147.59±15.37 个 vs 114.56±10.57 个) 以及侵袭数量(111.83±12.58 个 vs 79.56±8.48个)，Beclin1(0.94±0.11 vs 0.62±0.07)，LC3-II/I 蛋白(1.27±0.13 vs 0.98±0.11) 水平均升高（q=4.171 ～ 6.012， 均P<0.05），凋亡率（4.56%±0.86% vs 8.67%±1.46%），FOXO3（0.17±0.01 vs 0.33±0.04），FOXM1（0.46±0.03 vs 0.70±0.08）蛋白水平降低（q=5.941，9.505，6.881，均P<0.05），差异具有统计学意义。JY-2 逆转了BUT 对MG-63/CDDP 细胞活性和化疗耐药性的有利影响。结论 BUT 可能通过激活FOXO3-FOXM1 信号通路调节骨肉瘤细胞的细胞活性和CDDP耐药性。
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.

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备注/Memo

备注/Memo:
基金项目：湖北省卫生健康委科研项目（WJ2023F090）。
作者简介：奥婷婷（1988-），女，硕士研究生，主治医师，研究方向：麻醉，E-mail：a2q97bw@163.com。
通讯作者：姜祯珍（1984-），女，本科，主治医师，研究方向：麻醉，E-mail：jiangzz3884@163.com。
更新日期/Last Update: 2024-11-15