How Cancer Cells Escape Ferroptosis [July 9, 2025] DOJINDO LABORATORIES

Ferroptosis, a regulated and non-apoptotic form of cell death, has gained increasing attention in cancer and neuroscience, particularly as a therapeutic strategy against apoptosis-resistant tumors in cancer research¹. However, cancer cells can develop resistance. Recent studies have identified 7-dehydrocholesterol as an endogenous suppressor of ferroptosis that blocks lipid peroxidation². Elucidating these resistance mechanisms and promoting phospholipid peroxidation may lead to novel approaches in cancer therapy.

1. The cell biology of ferroptosis (Nature Reviews Molecular Cell Biology, 2024)
Summary: This review outlines the general mechanisms of ferroptosis and how lipid peroxides induce membrane rupture without pore-forming proteins, distinguishing it from other forms of cell death. It also highlights the roles of cellular organelles in ferroptosis, particularly the ER and lipid droplets involved in lipid metabolism. As PUFA-rich phospholipids in the cell membrane are highly susceptible to peroxidation, their synthesis and incorporation into membranes are key determinants of ferroptosis sensitivity. This review clarifies which cellular events researchers should focus on, making it a practical resource for future studies.

Related technique Lipid Peroxide Detection, Lipid Droplet Detection

2. 7-Dehydrocholesterol dictates ferroptosis sensitivity (Nature, 2024)
Summary: This study identifies 7-dehydrocholesterol (7-DHC) as a natural suppressor of ferroptosis, acting through inhibition of phospholipid peroxidation. By manipulating key enzymes in cholesterol biosynthesis, the authors demonstrate that altering 7-DHC levels can modulate ferroptosis sensitivity in cancer and ischemia–reperfusion injury models, suggesting potential therapeutic applications.

Highlighted technique: Using genome-wide CRISPR–Cas9 screening and targeted gene knockouts, this study identified cholesterol biosynthesis enzymes that modulate ferroptosis sensitivity through regulation of 7-dehydrocholesterol (7-DHC) levels. The anti-ferroptotic role of 7-DHC was confirmed by quantifying lipid peroxidation using fluorescent probes.

Related technique Intracellular Fe²⁺ Detection (Used in this article), Mitochondrial Lipid Peroxide Detection (Used in this article)

Related Techniques (click to open/close)

Target	Kit & Probes
Ferroptosis Indicator: ferrous ion (Fe²⁺)	FerroOrange(intracellular), Mito-FerroGreen(mitochondria)
Ferroptosis Indicator: lipid peroxidation	Liperfluo(intracellular), MitoPeDPP(mitochondria)
Lipid Peroxidation Assay	Lipid Peroxidation Probe -BDP 581/591 C11-
Cystine Uptake detection	Cystine Uptake Assay Kit
Glutathione Quantification	GSSG/GSH Quantification Kit
Mitochondrial superoxide detection	MitoBright ROS Deep Red - Mitochondrial Superoxide Detection
Total ROS detection	Highly sensitive DCFH-DA or Photo-oxidation Resistant DCFH-DA
Cell proliferation/ cytotoxicity assay	Cell Counting Kit-8 and Cytotoxicity LDH Assay Kit-WST

Application Note (click to open/close)
> When Lysosomes Go Neutral: Iron Loss Unveiled

We investigated the transition of cellular metabolisms in A549 cells treated with erastin, a known ferroptosis inducer. Our results revealed the following.

Results
- The inhibition of cystine uptake by erastin led to a depletion of cysteine, which in turn increased the compensatory uptake of other amino acids.
- Glucose uptake, which typically promotes ferroptosis*, was found to decrease upon erastin treatment, suggesting a potential cellular self-defense mechanism.
- The depletion of cysteine resulted in a decrease in glutathione levels and an increase in Fe²⁺, ROS, and lipid peroxides, all of which are recognized markers of ferroptosis.

Cell Line: A549
Incubation Conditions: 100 μmol/l Erastin/MEM, 37℃, 3h
*Reference: Xinxin Song, et al., Cell Reports, (2021)

Products in Use
① Amino Acid Uptake: Amino Acid Uptake Assay Kit
② Glucose Uptake: Glucose Uptake Assay Kit-Green
③ Cystine Uptake: Cystine Uptake Assay Kit
④ Intracellular glutathione: GSSG/GSH Quantification Kit
⑤ Intracellular labile Fe: FerroOrange
⑥ Intracellular total ROS: ROS Assay Kit -Highly Sensitive DCFH-DA-
⑦ Lipid Peroxides: Liperfluo