氯喹对博来霉素和奈达铂诱导的舌鳞癌SCC25细胞凋亡的影响

Ruoyu Wang, Lan Yang
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摘要

目的:探讨氯喹诱导化疗药物博来霉素和奈达铂对舌鳞癌SCC25细胞凋亡的影响及其机制。方法:采用MTT法筛选不同药物对SCC25细胞生长的抑制作用。采用MTT、Hoechst33258染色及流式细胞仪观察不同处理后SCC25细胞形态、细胞周期及凋亡的变化。化疗药物对SCC25细胞Bax、Bcl-2和NFkB凋亡相关蛋白表达的影响结果:氯喹对博来霉素和奈达铂诱导的SCC25细胞增殖抑制率显著高于博来霉素和奈达铂单用组(p<0.05);Hoechst33358荧光染色及流式细胞术显示,在博来霉素组中,氯喹+博来霉素组细胞凋亡率显著升高,且氯喹+奈达铂组细胞凋亡率显著高于奈达铂组(p<0.05)。氯喹+博来霉素组G0/G1期细胞比例高于博来霉素组(p<0.05), S期细胞与G2/M期细胞比例降低;同一氯喹+奈达铂组G0/G1期细胞比例高于奈达铂组(p<0.05), S期和G2/M期细胞比例下降。WB结果显示,氯喹+博来霉素组Bax和NFkB的表达水平明显高于博来霉素单独组,Bcl-2的表达水平明显低于博来霉素单独组。氯喹+奈达铂组Bax和NFkB的表达水平明显高于单用奈达铂组。Bcl-2表达水平低于单用奈达铂组,差异有统计学意义(p<0.05)。结论:氯喹诱导的博来霉素和奈达铂能显著抑制舌鳞癌SCC25细胞的增殖,其促凋亡作用可能与激活线粒体凋亡相关蛋白和核转录因子有关。
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Effect of chloroquine on the apoptosis of tongue squamous cell carcinoma SCC25 cells induced by bleomycin and nedaplatin
Objective: To study the effect and mechanism of chloroquine-induced chemotherapy drugs bleomycin and nedaplatin on apoptosis of tongue squamous cell carcinoma SCC25 cells. Methods: The inhibitory effect of different drugs on the growth of SCC25 cells was screened by MTT assay. MTT, Hoechst33258 staining and FCM were used to observe the changes of morphology, cell cycle and apoptosis of SCC25 cells after different treatments. The quantitative analysis of chloroquine induced by Western blot The effect of chemotherapy drugs on the expression of Bax, Bcl-2 and NFkB apoptosis- related proteins in SCC25 cells. Results: The proliferation inhibition rate of bleomycin and nedaplatin SCC25 cells induced by chloroquine was significantly higher than that of bleomycin and nedaplatin alone (p<0.05); Hoechst33358 fluorescence staining and flow cytometry showed In the bleomycin group, the apoptotic rate of the chloroquine plus bleomycin group was significantly increased, and the apoptosis rate of the chloroquine + nedaplatin group was significantly higher than that of the nedaplatin group (p<0.05). The proportion of G0/G1 phase in the chloroquine + bleomycin group was higher than that in the bleomycin group (p<0.05), and the ratio of cells in the S phase to the G2/M phase was decreased; the same chloroquine + nedaplatin group G0/G1 The proportion of cells in the period was higher than that in the nedaplatin group (p<0.05), and the proportion of cells in the S phase and G2/M phase decreased. WB results showed that the expression levels of Bax and NFkB in chloroquine + bleomycin group were significantly higher than those in bleomycin group alone, and the expression level of Bcl-2 was lower than that in bleomycin group alone. Bax and NFkB in chloroquine + nedaplatin group The expression level was significantly higher than that of the nedaplatin group alone. The expression level of Bcl-2 was lower than that of the nedaplatin group alone, and the difference was significant (p<0.05). Conclusion: Chloroquine induced bleomycin and nedaplatin can significantly inhibit the proliferation of tongue squamous cell carcinoma SCC25 cells, and its proapoptotic effect may be related to the activation of mitochondrial apoptosis-related proteins and nuclear transcription factors.
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