Licochalcone D Induces ROS-Dependent Apoptosis in Gefitinib-Sensitive or Resistant Lung Cancer Cells by Targeting EGFR and MET

Abstract

Licochalcone D (LCD), a flavonoid isolated from a Chinese medicinal plant Glycyrrhiza inflata, has a variety of pharmacological activities. However, the anti-cancer effects of LCD on non-small cell lung cancer (NSCLC) have not been investigated yet. The amplification of MET (hepatocyte growth factor receptor) compensates for the inhibition of epidermal growth factor receptor (EGFR) activity due to tyrosine kinase inhibitor (TKI), leading to TKI resistance. Therefore, EGFR and MET can be attractive targets for lung cancer. We investigated the anti-proliferative and apoptotic effects of LCD in lung cancer cells HCC827 (gefitinib-sensitive) and HCC827GR (gefitinib-resistant) through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, pull-down/kinase assay, cell cycle analysis, Annexin-V/7-ADD staining, reactive oxygen species (ROS) assay, mitochondrial membrane potential (MMP) assay, multi-caspase assay, and Western blot analysis. The results showed that LCD inhibited phosphorylation and the kinase activity of EGFR and MET. In addition, the predicted pose of LCD was competitively located at the ATP binding site. LCD suppressed lung cancer cells growth by blocking cell cycle progression at the G2/M transition and inducing apoptosis. LCD also induced caspases activation and poly (ADP-ribose) polymerase (PARP) cleavage, thus displaying features of apoptotic signals. These results provide evidence that LCD has anti-tumor effects by inhibiting EGFR and MET activities and inducing ROS-dependent apoptosis in NSCLC, suggesting that LCD has the potential to treat lung cancer.

Transcriptome Profiling and Cytological Assessments for Identifying Regulatory Pathways Associated With Diorcinol N-Induced Autophagy in A3 Cells

“…Pyknotic karyomorphism has been found during autophagy in other leukemia cells and is induced by treatment with different kinds of anticancer drugs (i.e., 786-O cells and L-02 cells) ( Yuan et al., 2017 ; Chen et al., 2018 ). The fidelity of replication is affected by abnormal karyomorphism, and low fidelity of replication could lead to cell cycle arrest ( Dyshlovoy et al., 2020 ; Oh et al., 2020 ). Our findings indicate that DN arrested the cell cycle of A3 cells at the G1/S phase, which is similar to the effects of previously reported anticancer agents, such as triacanthine and licoricidin ( Ji et al., 2017 ; Shin et al., 2019 ).…”

Perspectives and controversies regarding the use of natural products for the treatment of lung cancer

“…In addition to triterpenes, about 300 polyphenols, including phenolic acids, flavonoids, flavanes, chalcones, and isoflavones, with a concentration of 1%–5% each, have been isolated from licorice root and licorice extract 37 . Among them, licorice chalcone A (LICA), Licochalcone B (LCB), and Licochalcone D (LCD) have been proven to provide a variety of health benefits, including anticancer effects 38‐41 …”

“…Chiu et al also found that tanshinone IIA induces the apoptosis of A549 cells by inducing the release of reactive oxygen species and decreasing the MMP 129 . LCD can induce apoptosis through the accumulation of ROS in lung cancer cells, increasing the loss of MMP and the upregulation or downregulation of mitochondrial‐related proteins, which leads to apoptotic cell death through the mitochondrial (intrinsic) pathway 41 . To amplify the therapeutic effect of EGCG, magnetic nanoparticles (MNP) encapsulated in bovine serum albumin (BSA) and loaded with EGCG were synthesized to induce apoptosis through the participation of ROS, resulting in the loss of MMP, increase in the expression of Nrf2 and Keap1, regulation of the apoptosis of A549 cells, and induction of EGCG to exert a more substantial anti‐lung cancer effect 130 .…”

Natural products and phytochemical nanoformulations targeting mitochondria in oncotherapy: an updated review on resveratrol

“…Overall, triggering reactive oxygen species (ROS) production is a promising strategy in cancer therapy, since these reactive species are involved in stimulation of apoptosis through mitochondrial dysfucntion and much attention has been directed toward enhancing the ROS generation in cancer cells to suppress their malignancy and proliferation [66,67]. To date, there has been no article to investigate the relationship between the inhibitory effect of resveratrol on ROS production and its anti-tumor activity.…”