Ammonium Ferric Citrate induced Ferroptosis in Non-Small-Cell Lung Carcinoma through the inhibition of GPX4-GSS/GSR-GGT axis activity

Abstract

The morbidity and mortality rates associated with non-small-cell lung carcinoma (NSCLC) are increasing every year, placing new demands on existing therapies and drugs. Ammonium ferric citrate (AFC) is often used as a food additive for iron supplementation; however, to our knowledge, no studies have investigated whether AFC can induce ferroptosis in NSCLC. In this study, we demonstrated that specific concentrations of AFC effectively inhibit the proliferation and invasion of lung cancer cell lines in vitro using a cell proliferation inhibition test, a transwell assay, and flow cytometry analysis of cell cycle and apoptosis. In addition, AFC significantly induced oxidative stress injury in lung cancer cell lines. A quantitative polymerase chain reaction assay showed that AFC markedly reduced the expression levels of cell growth factors, negative regulators of ferroptosis, and autophagy regulators. Lastly, a protein-protein interaction analysis revealed that glutathione peroxidase 4 (GPX4) exerted its biological role through the regulation of the GSS/GSR complex and downstream GGT family proteins. When the expression of GPX4 changes, its biological activities, such as the glutathione metabolic process, cellular biosynthetic process, cellular response to chemical stimulus, and antioxidant activity, change accordingly, thereby affecting the survival quality and physiological and biochemical activities of cells. Overall, this study verifies that AFC has the biological activity of activating oxidative stress injury in NSCLC cell lines, leading to a decrease in their autophagy and inducing ferroptosis. We also confirmed that the GPX4-GSS/GSR-GGT axis is a crucial target of AFC-induced ferroptosis.

CircDTL Functions as an Oncogene and Regulates Both Apoptosis and Ferroptosis in Non-small Cell Lung Cancer Cells

“…These findings suggested that inhibition of circDTL induced apoptosis via the intrinsic pathway. Considering that most chemotherapeutic agents primarily induce apoptosis via the intrinsic route ( Wu et al, 2021 ), it might explain why the silencing of circDTL increased the chemosensitivity of NSCLC cells. Ferroptosis is a novel identified form of cell death.…”

“…For example, blockage of GPX4 could overcome resistance to Lapatinib via promoting ferroptosis in NSCLC (Ni et al, 2021). Another study also found that ammonium ferric citrate induced ferroptosis via blockage of GPX4 in NSCLC cells (Wu et al, 2021). Interestingly, several researchers have found that non-coding RNAs also play essential roles in regulating ferroptosis (Wang et al, 2019;Lu et al, 2020).…”

A Ferroptosis‐Related Prognostic Risk Score Model to Predict Clinical Significance and Immunogenic Characteristics in Glioblastoma Multiforme

“…Interactions between the rapamycin kinase and GPX4 targets may regulate autophagy-dependent ferroptosis in cancer cells. GPX4 downregulation enhances sensitivity to chemotherapy by promoting ferroptotic cell death [ 42 , 43 ]. Lipid peroxidation is positively regulated by ALOX5 and contributes to ferroptotic cell death.…”

Dexmedetomidine is Resistant to Ferroptosis by Activating mTOR-TFR1 Signaling

“…According to the literature [2,17,18,19], we found that the most appropriate FAC concentration for HT22 cells was 150 µM and incubated for 24h. This concentration was used as the stimulus concentration for subsequent experiments.…”

“…3B) in HT22 cells. The characteristic of mitochondria in Ferroptosis is that the mitochondrial membrane thickens and the membrane density increases, which causes the mitochondria to depolarize [18], ferroptosis is also accompanied by increased autophagosomes [19]. We carried out the detection of intracellular ultrastructure (mitochondria and nucleus).…”