ERK Activation-Mediated Autophagy Induction Resists Licochalcone A-Induced Anticancer Activities in Lung Cancer Cells in vitro.
ERK Activation-Mediated Autophagy Induction Resists Licochalcone A-Induced Anticancer Activities in Lung Cancer Cells in vitro.
复制标题
ERK 激活介导的自噬诱导可抵抗甘草查尔酮 A 诱导的肺癌细胞体外抗癌活性
DOI：
10.2147/ott.s278268
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发表时间：
2020
影响因子：
4
通讯作者：
Wen H
中科院分区：
文献类型：
--
作者：
Luo W;Sun R;Chen X;Li J;Jiang J;He Y;Shi S;Wen H
引言肺癌的发病率和死亡率在我国各类癌症中均居首位。 Licochalcone A (LA) 是从甘草根中提取的黄酮类化合物，在体外和体内对多种癌症具有抗肿瘤作用。然而，LA 在非小细胞肺癌 (NSCLC) 中的作用仍不清楚。方法采用MTT法、Edu染色法和集落形成法测定细胞活力。使用膜联蛋白 V/PI 双染分析和流式细胞术研究细胞凋亡。采用实时定量RT-PCR检测相关蛋白mRNA的表达情况。 Western blotting用于研究相关蛋白的表达。结果结果显示，LA以剂量依赖性方式抑制NSCLC细胞增殖并诱导细胞凋亡。此外，LA 显着抑制 c-IAP1、c-IAP2、XIAP、Survivin、c-FLIPL 和 RIP1 的表达，而不影响 mRNA 水平。放线菌酮追逐试验表明，LA 大大降低了 Survivin、XIAP 和 RIP1 的稳定性。机理研究表明，LA 诱导细胞保护性自噬，因为用 CQ 阻断自噬可大大增强 LA 诱导的抗癌活性。此外，LA 在 A549 和 H460 细胞中以时间依赖性方式快速诱导 ERK 和 p38 激活，但抑制 c-Jun N 末端激酶 (JNK) 的活性；用抑制剂抑制 ERK 而不是 p38 可减弱 LA 诱导的自噬，同时显着增强 LA 诱导的肺癌细胞细胞毒性，并进一步促进凋亡相关蛋白的降解。讨论 本研究的结果为细胞凋亡相关蛋白和 MAPKs 通路在 LA 抗癌活性中的作用提供了新的见解。
Introduction The incidence and mortality rates of lung cancer rank top in the different types of cancers in China. Licochalcone A (LA) is a flavonoid extracted from the roots of licorice with antitumor effects in various cancers in vitro and in vivo. However, the role of LA in non-small cell lung cancer (NSCLC) remains largely unclear. Methods The cell viability was measured by MTT assay, Edu staining and colony formation assay. Apoptosis was investigated using Annexin V/PI double-stained assays with flow cytometry. Real-time quantitative RT-PCR was carried out to investigate the expression of mRNA of related proteins. Western blotting was used to investigate the expression of related proteins. Results The results show that LA inhibits the proliferation of NSCLC cells in a dose-dependent manner and induces apoptotic cell death. Moreover, LA significantly suppresses the expression of c-IAP1, c-IAP2, XIAP, Survivin, c-FLIPL and RIP1 without influencing the level of mRNA. Cycloheximide chase assay demonstrates that LA greatly decreases the stability of Survivin, XIAP and RIP1. Mechanistic studies indicate that LA induces cytoprotective autophagy since block of autophagy with CQ greatly enhances LA-induced anticancer activities. Furthermore, LA rapidly induces ERK and p38 activation in a time-dependent manner in both A549 and H460 cells, but suppresses the activities of c-Jun N-terminal kinase (JNK); suppression of ERK not p38 with inhibitor attenuates LA-induced autophagy, while it remarkably enhances LA-induced cytotoxicity in lung cancer cells and further promotes the degradation of apoptosis-related proteins. Discussion The results of this study provide novel insights on the role of apoptosis-related proteins and the MAPKs pathway in the anticancer activities of LA.