Apigenin Combined With Gefitinib Blocks Autophagy Flux and Induces Apoptotic Cell Death Through Inhibition of HIF-1α, c-Myc, p-EGFR, and Glucose Metabolism in EGFR L858R+T790M-Mutated H1975 Cells

Abstract

Cancer cells are characterized by abnormally increased glucose uptake and active bio-energy and biosynthesis to support the proliferation, metastasis, and drug resistant survival. We examined the therapeutic value of the combination of apigenin (a natural small-molecule inhibitor of Glut1 belonging to the flavonoid family) and gefitinib on epidermal growth factor receptor (EGFR)-resistant mutant non-small cell lung cancer, to notably damage glucose utilization and thus suppress cell growth and malignant behavior. Here, we demonstrate that apigenin combined with gefitinib inhibits multiple oncogenic drivers such as c-Myc, HIF-1α, and EGFR, reduces Gluts and MCT1 protein expression, and inactivates the 5′ adenosine monophosphate-activated protein kinase (AMPK) signaling, which regulates glucose uptake and maintains energy metabolism, leading to impaired energy utilization in EGFR L858R-T790M-mutated H1975 lung cancer cells. H1975 cells exhibit dysregulated metabolism and apoptotic cell death following treatment with apigenin + gefitinib. Therefore, the combined apigenin + gefitinib treatment presents an attractive strategy as alternative treatment for the acquired resistance to EGFR-TKIs in NSCLC.

Molecular Crosstalk Between MYC and HIF in Cancer

“…For example, combination of apigenin and gefitinib, an epidermal growth factor receptor (EGFR) inhibitor, to treat EGFR-resistant mutant non-small cell lung cancers impairs energy utilization and suppresses cell growth and malignant behavior. They inhibit the activity of several oncogenic drivers such as MYC, HIF1α, and EGFR, reduce the protein expression of Gluts and MCT1, and inactivate the 5 adenosine monophosphate-activated protein kinase (AMPK) signaling (Chen Z. et al, 2019). Together, targeting these downstream signaling pathways controlled by MYC-HIF crosstalk (Figure 2E) or directly co-targeting MYC and HIFs could emerge as effective therapeutics in advanced human cancers such as advanced ccRCCs.…”

JNK and Autophagy Independently Contributed to Cytotoxicity of Arsenite combined With Tetrandrine via Modulating Cell Cycle Progression in Human Breast Cancer Cells

“…In addition to the induction of apoptotic/necrotic cell death, our results also demonstrated that autophagic cell death and Sphase arrest ( Supplementary Figures 1 and 2) contributed to the cytocidal effects of the combined regimen of 10 mM As III +6.4 mM Tetra in MDA-MB-231 cells. Although autophagy has been linked to apoptotic/necrotic cell death in many cases (Nikoletopoulou et al, 2013;Yoshida, 2017;Chen et al, 2019), our experimental results demonstrated that the addition of either 3-MA or wortmannin, two autophagy inhibitors, successfully corrected the combined regimen-triggered S-phase arrest, however, had little effect on the apoptosis/necrosis induction (Figures 4 and 5). Autophagy has been demonstrated to act either as a cytoprotective process or a pro-death factor in different cellular contexts (White and DiPaola, 2009; After treatment with 10 mM As III +6.4 mM Tetra in the presence or absence of 3-MA (0.25, 1.0 mM) or wortmannin (0.25, 1.0 mM) for 48 h, cell cycle profiling was performed by FACSCanto flow cytometer.…”

“…Herein, both the induction of autophagy and S-phase arrest were first confirmed in MDA-MB-231 cells following the exposure to the combined regimen of 10 mM As III +6.4 mM Tetra for 48 h ( Supplementary Figures 1 and 2). Previous studies have demonstrated a close association between autophagy and apoptosis as well as cell cycle arrest induction in different types of cancer cells including MDA-MB-231 (Cheng et al, 2015;Gao et al, 2015;Lee et al, 2016;Lin et al, 2016;Chen et al, 2019). In order to clarify whether there was a link between autophagy and apoptosis/necrosis as well as S-phase arrest, two autophagy inhibitors, 3-MA and wortmannin, were used in the current study.…”

Effect of apigenin on whole transcriptome profile of TNFα‑activated MDA‑MB‑468 triple negative breast cancer cells

“…Overall, we believe there is enough support to warrant clinical trials for the use of apigenin, as there is a growing body of researching showing its antitumor effects from multiple stand points from blocking mutagenic induced cancers [e.g., methyl-nitrosourea, methyl-n-nitro-N-nitrosoguanidine, benzo(a)pyrene or 2-aminoanthracene] (88) to inhibition of ornithine decarboxylase (89) and its overall antioxidant, anti-inflammatory effects (90,91). Data on the clinical efficacy of substances like apigenin for human use to reduce CLL2 will also need to be confirmed, as well as establishing its bioavailability, absorption, therapeutic concentration and application (prevention, treatment or for chemotherapy drug augmentation) (55,(91)(92)(93)(94).…”