Eriocitrin inhibits epithelial-mesenchymal transformation (EMT) in lung adenocarcinoma cells via triggering ferroptosis

Abstract

Introduction: Lung adenocarcinoma (LUAD) is the most prevalent pathological subtype of non-small cell lung cancer (NSCLC), characterized by a high propensity for relapse and metastasis due to epithelial-mesenchymal transition (EMT) of cancer cells. Ferroptosis, a newly discovered regulated cell death modality, is interconnected with the EMT process in certain cancers. Eriocitrin, a natural flavonoid compound, exerts anti-inflammatory and anticancer effects. Objectives: The aim of this study is to investigate the potential inhibitory effect of eriocitrin on lung adenocarcinoma metastasis and explore whether its underlying mechanism involves ferroptosis induction in cancer cells. Methods: The CCK8 assay and wound healing assay and transwell were conducted to determine the cell viability and migration ability of A549 and H1299 cells, respectively. EMT process was assessed by western blot and RT-PCR to detect protein and mRNA levels of EMT markers. ROS and cell iron were measured to determine ferroptosis level. Results: Eriocitrin treatment significantly inhibited cell viability and migration ability in a concentration-dependent manner. Furthermore, eriocitrin administration for 24 hours resulted in enhanced expression of E-cadherin, while downregulating vimentin, N-cadherin and snail expression, indicating marked repression of the EMT process. Additionally, eriocitrin significantly induced ferroptosis in A549 and H1299 cells, as evidenced by increased ROS levels, downregulation of Nrf-2, SLC7A11 and GPX4 expression, and enhanced cellular iron accumulation. Moreover, pretreatment with the ferroptosis inhibitor ferrostatin-1 effectively abrogated the inhibitory effects of eriocitrin on EMT. Conclusions: Our findings further support the anti-cancer properties of eriocitrin, as evidenced by its ability to inhibit the EMT process in LUAD cells, which is partially mediated through induction of ferroptosis in cancer cells.

Eriocitrin Inhibits Angiogenesis by Targeting VEGFR2-Mediated PI3K/AKT/mTOR Signaling Pathways

“…Our experimental results have shown that in vitro angiogenesis can be significantly inhibited by non-toxic concentrations of eriocitrin. Previous studies have shown that eriocitrin can inhibit cancer movement by inhibiting cell movement during the wound healing process in A549 and H1299 cancer cells [9]. Our results also suggest that eriocitrin can help prevent tumor growth by inhibiting capillary formation and migration during the initial phases of angiogenesis.…”

“…Eriocitrin, also known as eriodictyol 7-rutinoside, is a flavanone compound abundant in peppermint, lemon, and various citrus fruits [6,7]. Eriocitrin exhibits potent biological effects, including its strong antioxidant [8], antitumor [9,10], anti-allergic [11], and antiinflammatory properties [12][13][14]. Recent studies have shown that eriocitrin can activate pro-apoptotic proteins such as Bax, caspases-7, -8, and -9, while inhibiting the expression of cytoprotective proteins Bcl-2 and Bcl-x in MCF-7 cells [15].…”

Eriocitrin Disrupts Erythrocyte Membrane Asymmetry through Oxidative Stress and Calcium Signaling and the Activation of Casein Kinase 1α and Rac1 GTPase

“…The current study reveals, for the first time, that anticancer concentrations of ERN (20-100 µM) [12][13][14][15] stimulate premature RBC death by both hemolysis and eryptosis. However, whether this concentration range is achievable in vivo remains to be determined in future studies.…”

“…Moreover, ERN promotes intrinsic apoptosis in MCF-7 cells through JAK2/STAT3/Src inhibition and reactive oxygen species (ROS)-dependent JNK/p38 MAPK stimulation [14]. Very recently, ERN has been reported to trigger ferroptosis in A549 and H1299 lung adenocarcinoma cells [15], thereby circumventing cancer cell resistance to apoptosis. Altogether, mounting studies are arguing for the further development of ERN as an anticancer therapeutic.…”

The role of lipid peroxidation in epithelial–mesenchymal transition of retinal pigment epithelial cells

“…Similarly, in lung adenocarcinoma cells (A549 and H1299), an antioxidant eriocitrin suppressed EMT, upregulated ROS levels, and induced ferroptosis. Fer-1 abrogated the inhibitory effects of eriocitrin on EMT 32 . Another study using A549 cells have also shown that Erastin-induced lipid peroxidation could be delayed, by TGF-β1-induced EMT 33 .…”