Department of Pharmacy, College of Pharmacy, Mokpo National University, Jeonnam 58554, Republic of Korea.
China-US (Henan) Hormel Cancer Institute, Zhengzhou, Henan 450008, PR China; Basic Medical College, Zhengzhou University, Zhengzhou, Henan 450001, PR China.
Phytomedicine. 2019 Oct;63:153014. doi: 10.1016/j.phymed.2019.153014. Epub 2019 Jul 5.
Epidermal growth factor receptor (EGFR) gene alterations are associated with sensitization to tyrosine kinase inhibitors such as gefitinib in lung cancer. Some patients suffering from non-small cell lung cancer (NSCLC) have difficulty in treating the cancer due to resistance acquired to gefitinib with MET amplification. Therefore EGFR and MET may be attractive targets for lung cancer therapy.
This study aimed to investigate the anti-cancer activity of Licochalcone (LC)B extracted from Glycyrrhiza inflata, in gefitinib-sensitive or gefitinib-resistant NSCLC cells, and to define its mechanisms.
We investigated the mechanism of action of LCB by targeting EGFR and MET in human NSCLC cells.
We used the HCC827 and HCC827GR lines as gefitinib-sensitive and -resistant cells respectively, and determined the effects of LCB on both, by performing cell proliferation assay, flow cytometry analysis and Western blotting. Targets of LCB were identified by pull-down/kinase assay and molecular docking simulation.
LCB inhibited both EGFR and MET kinase activity by directly binding to their ATP-binding pockets. The ability of this interaction was verified by computational docking and molecular dynamics simulations. LCB suppressed viability and colony formation of both HCC827 and HCC827GR cells while exhibiting no cytotoxicity to normal cells. The induction of G2/M cell-cycle arrest and apoptosis by LCB was confirmed by Annexin V/7-AAD double staining, ER stress and reactive oxygen species induction, mitochondrial membrane potential loss and caspase activation as well as related-proteins regulation. Inhibition of EGFR and MET by LCB decreased ERBB3 and AKT axis activation.
We provide insights into the LCB-mediated mechanisms involved in reducing cell proliferation and inducing apoptosis in NSCLC cells. This occurs through dual inhibition of EGFR and MET in NSCLC cells regardless of their sensitivity or resistance to gefitinib. LCB may be a promising novel therapeutic medicine for gefitinib-sensitive or resistant NSCLC treatment.
表皮生长因子受体（EGFR）基因改变与肺癌对酪氨酸激酶抑制剂（如吉非替尼）的敏感性有关。一些患有非小细胞肺癌（NSCLC）的患者由于 MET 扩增导致对吉非替尼产生耐药性，因此难以治疗癌症。因此，EGFR 和 MET 可能成为肺癌治疗的有吸引力的靶点。
本研究旨在探讨从甘草中提取的甘草查尔酮 B（LCB）在吉非替尼敏感或耐药的 NSCLC 细胞中的抗癌活性，并确定其机制。
我们通过靶向人 NSCLC 细胞中的 EGFR 和 MET 来研究 LCB 的作用机制。
我们分别使用 HCC827 和 HCC827GR 细胞作为吉非替尼敏感和耐药细胞，并通过细胞增殖测定、流式细胞术分析和 Western blot 来确定 LCB 对两者的影响。通过下拉/激酶测定和分子对接模拟确定 LCB 的靶点。
LCB 通过直接结合其 ATP 结合口袋抑制 EGFR 和 MET 激酶活性。这种相互作用的能力通过计算对接和分子动力学模拟得到验证。LCB 抑制 HCC827 和 HCC827GR 细胞的活力和集落形成，而对正常细胞无细胞毒性。通过 Annexin V/7-AAD 双重染色、内质网应激和活性氧诱导、线粒体膜电位丧失和半胱天冬酶激活以及相关蛋白调节，证实了 LCB 诱导 G2/M 细胞周期停滞和细胞凋亡。LCB 抑制 EGFR 和 MET 减少了 ERBB3 和 AKT 轴的激活。
我们提供了关于 LCB 介导的机制的见解，这些机制涉及减少 NSCLC 细胞的增殖并诱导其凋亡。这是通过在 NSCLC 细胞中双重抑制 EGFR 和 MET 来实现的，而与它们对吉非替尼的敏感性或耐药性无关。LCB 可能是一种有前途的新型治疗药物，可用于治疗吉非替尼敏感或耐药的 NSCLC。
