抑制内质网应激信号通路PERK-eIF2α介导NF-κB抑制口咽癌细胞增殖

摘要

Objective: To explore the mechanism of regulation proliferation in oropharyngeal cancer cells by endoplasmic reticulum stress signaling pathway PERK-eIF2α. Methods: MTT assay was used to detect proliferation in oropharyngeal cancer cells (Fadu, Detroit 562) by different concentrations (0 μmmol／L, 1 μmmol／L, 10 μmmol／L, 50 μmmol／L) of PERK pathway inhibitor GSK2606414 and NF-κB inhibitor Bay11-7082. By Western Blot assay, we observed the activation of PERK-eIF2α-NF-κB signaling pathway. Cell apoptosis rate was detected by flow. Results: The results of MTT assay showed that with the increase of pretreatment concentration of GSK2606414, the survival rate of Fadu cells and Detroit 562 cells gradually decreased (F=56. 06, P=0. 000, F=71. 13, P=0. 000). Secondly, silencing PERK induced apoptosis. We further explored its mechanism and found that silencing PERK inhibits NF-κB phosphorylation, suggesting that abnormal activation of PERK-eIF2α signaling pathway induces NF-κB phosphorylation. Finally, we inhibited NF-κB to verify the above results. The results of MTT assay showed that with the in-crease of pretreatment concentration of Bay11 7082, the survival rate of Fadu cells and Detroit 562 cells decreased gradually (F=57. 48, P=0. 000, F=116. 76, P=0. 000). The apoptosis rate of Fadu cells and Detroit 562 cells in Bay11-7082 group was significantly higher than that in the control group (t=12. 38, 20. 88; P=0. 007, 0. 002). All of them showed in-hibition of cell proliferation and induced apoptosis. Conclusion: Inhibition of endoplasmic reticulum stress signaling pathway PERK-eIF2α mediates NF-κB inhibition of oropharyngeal cancer cell proliferation.%目的:探讨内质网应激信号通路PERK-eIF2α参与调控口咽癌细胞增殖具体机制.方法:采用MTT实验分别检测不同浓度(0 μmmol／L、1 μmmol／L、10 μmmol／L、50 μmmol／L)PERK通路抑制剂GSK2606414和NF-κB抑制剂Bay11-7082对口咽鳞癌细胞(Fadu、Detroit 562)增殖的影响;应用Western Blot实验检测PERK-eIF2α-NF-κB通路活化情况;Annexin V、PI染色、流式细胞仪检测凋亡分数.结果:首先应用PERK抑制剂预处理细胞,MTT结果示,随着GSK2606414预处理浓度的增加,Fadu细胞和Detroit 562细胞的存活率均逐渐降低(F=56. 06,P=0. 000;F=71. 13, P=0. 000).其次沉默PERK诱导细胞凋亡.我们进一步探讨其机制,发现沉默 PERK抑制 NF-κB磷酸化,提示PERK-eIF2α信号通路异常活化诱导NF-κB磷酸化.最后我们抑制NF-κB验证上述结果,MTT结果示,随着Bay11 7082预处理浓度的增加,Fadu细胞和Detroit 562 细胞的存活率均逐渐降低( F=57. 48,P=0. 001;F=116. 76,P=0. 001);同时,Bay11-7082组Fadu细胞、Detroit 562细胞凋亡比例分别较各自的control组明显增加,差异有统计学意义(t=12. 38、20. 88,P=0. 007、0. 002);均显示抑制细胞增殖诱导凋亡.结论:抑制内质网应激信号通路 PERK-eIF2α介导NF-κB抑制口咽癌细胞增殖.