布托啡诺抑制TLR4/MyD88/NF-κB信号通路对卵巢癌细胞增殖、凋亡、迁移和侵袭的影响

Influences of butorphanol on the proliferation, apoptosis, migration and invasion of ovarian cancer cells by inhibiting TLR4/MyD88/NF-κB signaling pathway

目的 探讨布托啡诺抑制Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核因子-κB(NF-κB)信号通路对卵巢癌细胞增殖、凋亡、迁移和侵袭的影响。方法 实验分组1:(2.5、5.0、10、20、40、80、160、320)μg/mL布托啡诺处理人卵巢癌细胞A2780,计算细胞半数抑制浓度(IC_(50))。实验分组2:对照组(正常培养)、布托啡诺组(13.24μg/mL布托啡诺)、布托啡诺+LPS组(13.24μg/mL布托啡诺+1μg/mL LPS),CCK-8检测细胞增殖情况;流式细胞术检测细胞凋亡情况;Transwell检测细胞侵袭和迁移情况;蛋白质免疫印迹检测细胞中TLR4、MyD88、NF-κB蛋白表达情况。结果 细胞IC_(50)为13.24μg/mL。与对照组相比,布托啡诺组细胞OD450水平、迁移和侵袭数量、细胞中TLR4、MyD88、NF-κB蛋白水平降低(P<0.05),细胞凋亡率升高(P<0.05);与布托啡诺组相比,布托啡诺+LPS组细胞OD450水平、迁移和侵袭数量、细胞中TLR4、MyD88、NF-κB蛋白水平升高(P<0.05),细胞凋亡率降低(P<0.05)。结论 布托啡诺能够抑制TLR4/MyD88/NF-κB信号通路,进而抑制细胞增殖、迁移和侵袭,促进细胞凋亡。

Objective To investigate the influences of butorphanol on the proliferation, apoptosis, migration and invasion of ovarian cancer cells by inhibiting the Toll-like receptor 4(TLR4)/myeloid differentiation factor 88(MyD88)/nuclear factor-κB(NF-κB) signaling pathway. Methods Experimental group 1:(2.5, 5.0, 10, 20, 40, 80, 160, 320)μg/mL butorphanol was used to treat human ovarian cancer cells A2780, and the cell median inhibitory concentration(IC_(50))was calculated. Experimental group 2: control group(normal culture), butorphanol group(13.24 μg/mL butorphanol), and butorphanol+LPS group(13.24 μg/mL butorphanol+1 μg/mL LPS). CCK-8 was performed to determine cell proliferation.Flow cytometry was performed to determine apoptosis. Transwell was implemented to detect cell invasion and migration.Western blot was performed to determine TLR4, MyD88, NF-κB protein expression in cells. Results The cell IC_(50) was 13.24μg/mL. Compared with the control group, the level of OD450, the numbers of migration and invasion, the protein levels of TLR4, MyD88 and NF-κB in the butorphanol group were decreased(P<0.05), and the apoptosis rate was increased(P<0.05).Compared with the butorphanol group, the level of OD450, the numbers of migration and invasion, the protein levels of TLR4,MyD88 and NF-κB in the butorphanol+LPS group were increased(P<0.05), and the apoptosis rate was decreased(P<0.05).Conclusion Butorphanol can inhibit TLR4/MyD88/NF-κB signaling pathway, thereby inhibiting cell proliferation, migration and invasion, and promoting cell apoptosis.