Dual inhibition of EGFR and MET by Echinatin retards cell growth and induces apoptosis of lung cancer cells sensitive or resistant to gefitinib

Abstract

Patients with non-small-cell lung cancer (NSCLC) containing epidermal growth factor receptor (EGFR) amplification or sensitive mutations initially respond to tyrosine kinase inhibitor gefitinib; however, the treatment is less effective over time. Gefitinib resistance mechanisms include MET gene amplification. A therapeutic strategy targeting MET as well as EGFR can overcome resistance to gefitinib. In the present study we identified Echinatin (Ecn), a characteristic chalcone in licorice, which inhibited both EGFR and MET and strongly altered NSCLC cell growth. The antitumor efficacy of Ecn against gefitinib-sensitive or -resistant NSCLC cells with EGFR mutations and MET amplification was confirmed by suppressing cell proliferation and anchorageindependent colony growth. During the targeting of EGFR and MET, Ecn significantly blocked the kinase activity, which was validated with competitive ATP binding. Inhibition of EGFR and MET by Ecn decreases the phosphorylation of downstream target proteins ERBB3, AKT and ERK compared with total protein expression or control.Ecn induced the G2/M cell cycle arrest, and apoptosis via the intrinsic pathway of caspase-dependent activation. Ecn induced ROS production and GRP78, CHOP, DR5 and DR4 expression as well as depolarized the mitochondria membrane potential. Therefore, our results suggest that Ecn is a promising therapeutic agent in NSCLC therapy. KEYWORDS Echinatin, EGFR, MET, Non-small cell lung cancer, ApoptosisAbbreviations: 7-AAD, 7-aminoactinomycin D; Apaf-1, apoptotic protease activating factor-1; CHOP, CCAAT/enhancer-binding protein homologous protein; cyto c, cytochrome c; C-PARP, cleaved poly (ADP-ribose) polymerase; DMSO, dimethyl sulfoxide; DR, death receptor; Ecn, Echinatin; EGFR, epidermal growth factor receptor; FBS, fetal bovine serum; GRP78, 78-kDa glucose-regulated protein; MMP, mitochondrial membrane potential; MTT, 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide; NSCLC, Non-small cell lung cancer; PBS, phosphatebuffered saline; ROS, reactive oxygen species; RT, room temperature; RTK, receptor tyrosine kinase; TKI, tyrosine kinase inhibitor Ha-Na Oh and Mee-Hyun Lee equally contributed to this work.

Artemether, Artesunate, Arteannuin B, Echinatin, Licochalcone B and Andrographolide Effectively Inhibit SARS-CoV-2 and Related Viruses In Vitro

“…By screening the antiviral effects of 389 monomer compounds in the “three formulas and three medicines” (TargetMol, L6720, Supplementary Table S1 ) at the final concentration of 50 μM, six of the monomer compounds have been found to reduce the GX_P2V infection remarkably ( Figure 2A , Supplementary Figure S1 and Supplementary Table S2 ) . Among these six drugs, high concentrations of two anticancer drugs including echinatin ( Oh et al., 2020 ) (EC 50 = 20.89 µM, CC 50 = 120.1 µM, SI = 5.75), and licochalcone B ( Song et al., 2020 ) (EC 50 = 24.90 µM, CC 50 = 106.5 µM, SI = 4.28) were required to inhibit the viral infection ( Figure 2B ) . Notably, andrographolide ( Enmozhi et al., 2020 ) (EC 50 = 6.786 µM, CC 50 = 95.73 µM, SI = 14.11) were also found to block the virus infection ( Figure 2B ) .…”

Molecular Mechanisms of Antiproliferative Effects of Natural Chalcones

“…Additionally, the ability to induce the intrinsic pathway of apoptosis has also been described in several natural chacones such as echinatin [ 73 ], broussochalcone [ 74 ], hydroxysafflor yellow A [ 75 ], 3-deoxysappanchalcone [ 76 ], sappanchalcone [ 77 ], and many others (see also Table 1 ).…”

Echinatin mitigates H2O2-induced oxidative damage and apoptosis in lens epithelial cells via the Nrf2/HO-1 pathway

“…15 Studies have shown that Ech has a wide range of biological properties, including anti-inflammatory and anti-tumor effects. 16 Importantly, Liang et al 17 confirmed that Ech may undergo electron transfer and proton transfer to cause antioxidant effects. However, to date, the effects of Ech on oxidative stress-induced LEC damage have not been reported, and its molecular mechanism is largely unclear.…”