Erastin induces apoptotic and ferroptotic cell death by inducing ROS accumulation by causing mitochondrial dysfunction in gastric cancer cell HGC‑27

Abstract

erastin, a classical inducer of non-apoptotic cell death, exerts cytotoxicity in several types of cancer cells, including gastric cancer cells, by depleting glutathione, which is a primary cellular antioxidant, thus causing reactive oxygen species (roS) accumulation. although numerous studies have focused on the non-apoptotic cell death induced by erastin, whether erastin induces apoptosis remains unknown. The present study confirmed the cytotoxicity of erastin in HGC-27 cells and used a 30% inhibitory concentration (ic 30 , approximately 6.23 µM) for further analysis. The cell cycle analysis revealed that 6.23 µM of erastin inhibited proliferation by blocking the cell cycle at the G1/G0 phase. Further analysis also showed that 6.23 µM of erastin clearly inhibited HGC-27 malignant behaviors, including migration, invasion, colony formation and tumor formation in soft agar. The observation of roS accumulation due to erastin treatment led to determination of the effects of erastin on mitochondrial function and, as expected, erastin treatment decreased transcriptional activity and aTP production in mitochondria and disrupted the mitochondrial potential; these effects were reversed by the addition of the roS scavenger nac. To evaluate the effect of erastin in inducing apoptosis, HGC-27 cells were treated with 6.23 µM of erastin for 7 days and then analyzed. Evident apoptotic cell death was induced by erastin and this apoptosis was reversed by the addition of an apoptosis inhibitor (zVAD) or nac but not by the addition of a ferroptosis inhibitor (ferrostatin-1). Furthermore, the detection of caspase-3 and poly (adenosine diphosphate-ribose) polymerase (ParP) also confirmed that treatment with erastin promoted the cleavage of caspase-3 and ParP, which are hallmarks of apoptosis. Taken together, the present study revealed that a low dose of erastin inhibited malignant behavior and induced apoptosis by causing mitochondrial dysfunction.

A Cystine-Cysteine Intercellular Shuttle Prevents Ferroptosis in xCTKO Pancreatic Ductal Adenocarcinoma Cells

“…In our previous study [ 10 ], we showed that genetic disruption of the xCT transporter in pancreatic ductal adenocarcinoma (PDAC), one of the most aggressive types of cancer with a dismal 5-year survival rate of 7% [ 11 ], promotes intracellular cysteine depletion, inhibition of GSH synthesis followed by accumulation of membrane lipid hydroperoxides and finally, ferroptotic cell death in vitro. Similar results have been observed with pharmacological inhibition of xCT by erastin in a variety of cancers including breast, liver, gastric and pancreatic tumors [ 12 , 13 , 14 , 15 ]. However, in the same study our group demonstrated that, contrasting with the in vitro results, xCT-depleted PDAC cell xenografts in mice were able to grow at the same rate as their wild type (WT) counterparts, albeit with a slight delay.…”

Implications of Oxidative Stress in Glioblastoma Multiforme Following Treatment with Purine Derivatives

“…The induction of ferroptosis is mainly based on the Fenton reaction, wherein the accumulated iron (II) is oxidized into iron (III) by hydrogen peroxide, and lipid peroxides are in turn converted into ROS hydroxyl radical) [92]. Our results indicate that KR might be a potential inducer of ferroptosis and acts similar to erastin by inducing apoptotic cell death through ROS accumulation [94]. In contrast, Hu et al [93] demonstrated that ferroptosis might play an important role in the resistance of gliomas to temozolomide, which might be confirmed on the basis of the results obtained after 7-deazaKR treatment.…”

Sterculic Acid Alters Adhesion Molecules Expression and Extracellular Matrix Compounds to Regulate Migration of Lung Cancer Cells

“…Lipid peroxidation associated to ferroptosis could be an alternative cell-death mechanism, nevertheless Caspase activation has not been described in ferroptosis cell-death [41]. Simultaneous activation of some cell death mechanisms has been already observed [42,43]. We postulate that Caspase-3 activation could be the final step of an alternative/mixed cell-death mechanism, although pyroptosis could not be excluded as an alternative SA-induced cell-death mechanism.…”