【摘要】 目的探讨医用臭氧是否通过核因子(NF)-κB信号通路调控上皮-间质细胞转化(EMT)相关蛋白表达抑制宫颈癌细胞增殖和迁移。方法选用人宫颈癌Hela细胞,分别设空白组、N-乙酰-L-半胱氨酸(NAC)处理组、NAC和臭氧共处理组、臭氧组。细胞在臭氧处理前以60 mmol/mL NAC溶液预处理20 min,随后将细胞悬液与等体积半最大效应浓度(EC50)臭氧混合15 min。细胞计数试剂盒(CCK)-8和细胞集落实验检测细胞增殖抑制。二氯二乙酸酯(DCFH-DA)探针通过流式细胞术检测活性氧(ROS)水平。划痕实验和Transwell实验验检测臭氧处理后Hela细胞迁移能力。蛋白质印迹法检测NF-κB和EMT相关蛋白表达水平。结果臭氧处理后Hela细胞增殖能力下降,臭氧EC50为10 mg/mL,同时其迁移能力也下降。臭氧处理后,Hela细胞NF-κB和κB抑制性蛋白激酶(IKK)α表达及其磷酸化水平均降低,同时EMT相关波形蛋白(vimentin)和β-连环蛋白(β-catenin)表达水平降低,而NAC预处理可逆转臭氧的作用。结论医用臭氧可能通过抑制NF-κB信号通路下调vimentin、β-catenin抑制宫颈癌Hela细胞增殖和迁移,ROS在其中发挥重要作用,表明医用臭氧有望成为宫颈癌新的辅助治疗手段。更多还原

【Abstract】 Objective To investigate whether medical ozone inhibits the proliferation and migration of cervical cancer Hela cells by regulating epithelial-mesenchymal cell transformation-related(EMT-related) protein expressions through NF-κB signaling pathway. Methods Hela cells of human cervical cancer were used in this study. Blank group, N-acetyl-L-cysteine(NAC) treatment group, NAC and ozone combination treatment group and ozone treatment group were set up respectively. Before ozone treatment, the cells were pre-treated with 60 mmol/ml NAC solution for 20 min, then the cell suspension was mixed with equal volume of EC50 ozone for 15 min. CCK-8 kit and colony forming test were used to detect the cell proliferation inhibition. The level of reactive oxygen species(ROS) was detected by flow cytometry with a DCFH-DA probe. The migration ability of ozone-treated Hela cells was checked by scratch test and Transwell test. The expression levels of NF-κB and EMT-related proteins were detected by Western blot. Results After ozone treatment, the proliferation ability of Hela cells was decreased, the EC50 ozone was 10 mg/mL, besides, the migration ability of Hela cells was also decreased. The NF-κB and κB inhibitory protein kinase-α(IKK-α) expression and its phosphorylation level were decreased, in the same time the EMT-related vimentin and β-catenin expression levels were also decreased, and NAC pre-treatment could counter the effect of ozone. Conclusion Medical ozone can inhibit the proliferation and migration of cervical cancer Hela cells by down-regulating vimentin and β-catenin through inhibiting the NF-κB signaling pathway, in which ROS plays an important role, indicating that medical ozone may become a new adjuvant therapy for cervical cancer.(J Intervent Radiol, 2021, 30: 1140-1145)更多还原