【摘要】 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(0μmmol/L、1μmmol/L、10μmmol/L、50μmmol/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。更多还原

【Abstract】 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 increase 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 inhibition 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.更多还原