Mechanisms of Growth Inhibition by Sulfasalazine and Erastin in Hepatocellular Carcinoma Cell Lines

Kim, Do-Hyung; Abdullah,; Lee, Seung Jin

doi:10.17480/psk.2019.63.3.152

Cited by 3 publications

(9 citation statements)

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“…We evaluated the effects of necrostatin-1 on ferroptosis in Huh7 and SK-HEP-1 cells because we found these cell lines to be highly sensitive to ferroptosis in our previous study [ 17 ]. Treatment with sulfasalazine, erastin, and RSL3 for 24 h decreased cell viability in a dose-dependent manner.…”

Section: Resultsmentioning

confidence: 99%

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Necrostatin-1 Prevents Ferroptosis in a RIPK1- and IDO-Independent Manner in Hepatocellular Carcinoma

et al. 2021

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Ferroptosis is caused by the iron-mediated accumulation of lipid peroxidation, which is distinct from apoptosis and necroptosis. Necrostatin-1 inhibits receptor-interacting serine/threonine-protein kinase 1 (RIPK1) to initiate necroptosis; it also inhibits indoleamine 2,3-dioxygenase (IDO) to regulate tumor immunity. However, few studies have examined the off-target effect of necrostatin-1 on the ferroptosis pathway. The present study examined whether necrostatin-1 could interrupt ferroptosis induced by system xc- inhibitors (sulfasalazine and erastin) and a glutathione peroxidase 4 inhibitor (RSL3) in Huh7 and SK-HEP-1 cells. Necrostatin-1 completely prevented decreases in cell viability induced by sulfasalazine and erastin; it partially blunted decreases in cell viability induced by RSL3. Necrostatin-1, ferrostatin-1, and deferoxamine repressed sulfasalazine-provoked membrane permeabilization, as detected by 7-aminoactinomycin D staining and lipid peroxidation measured using a C11-BODIPY probe. However, other RIPK1 inhibitors (necrostatin-1s and GSK2982772) and an IDO inhibitor (1-methyl-D-tryptophan) did not recover the decrease in cell viability induced by sulfasalazine. Necrostatin-1 potentiated sulfasalazine-induced expression of xCT, a catalytic subunit of system xc- in these cells. These results demonstrated that necrostatin-1 blocked ferroptosis through a mechanism independent from RIPK1 and IDO inhibition in Huh7 and SK-HEP-1 cells, indicating that its antioxidant activity should be considered when using necrostatin-1 as a RIPK1 inhibitor.

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Section: Resultsmentioning

confidence: 99%

“…In our previous study, a sulfasalazine-induced decrease in cell viability was prevented by ferrostatin-1, although it was not prevented by Z-VAD-FMK and chloroquine [ 17 ]. Necrostatin-1 also had a protective effect against these cell deaths.…”

Section: Introductionmentioning

confidence: 99%

Necrostatin-1 Prevents Ferroptosis in a RIPK1- and IDO-Independent Manner in Hepatocellular Carcinoma

et al. 2021

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“…It is also overexpressed in several other cancers (46). It is commonly assumed that inhibition affects cells through decreased uptake of cystine (37,(47)(48)(49), which is thought to upset synthesis of GSH, rendering the cells vulnerable to oxidative stress. This is corroborated by observations that loss of viability at high inhibitor concentrations is rescued by addition of N-acetyl-cysteine or 2-mercaptoethanol (48,49), which enables uptake of cysteine by the cells.…”

Section: Discussionmentioning

confidence: 99%

“…It is commonly assumed that inhibition affects cells through decreased uptake of cystine (37,(47)(48)(49), which is thought to upset synthesis of GSH, rendering the cells vulnerable to oxidative stress. This is corroborated by observations that loss of viability at high inhibitor concentrations is rescued by addition of N-acetyl-cysteine or 2-mercaptoethanol (48,49), which enables uptake of cysteine by the cells. However, at low inhibitor concentrations (≤0.2 mM), such as the ones used in the present study, these studies show limited effect on viability, and it appears that strong inhibition is required to affect GSH levels to a degree where viability is compromised.…”

Section: Discussionmentioning

confidence: 99%

Quantitative analysis of amino acid metabolism in liver cancer links glutamate excretion to nucleotide synthesis

Nilsson

Haanstra

Engqvist

et al. 2020

Proc. Natl. Acad. Sci. U.S.A.

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Many cancer cells consume glutamine at high rates; counterintuitively, they simultaneously excrete glutamate, the first intermediate in glutamine metabolism. Glutamine consumption has been linked to replenishment of tricarboxylic acid cycle (TCA) intermediates and synthesis of adenosine triphosphate (ATP), but the reason for glutamate excretion is unclear. Here, we dynamically profile the uptake and excretion fluxes of a liver cancer cell line (HepG2) and use genome-scale metabolic modeling for in-depth analysis. We find that up to 30% of the glutamine is metabolized in the cytosol, primarily for nucleotide synthesis, producing cytosolic glutamate. We hypothesize that excreting glutamate helps the cell to increase the nucleotide synthesis rate to sustain growth. Indeed, we show experimentally that partial inhibition of glutamate excretion reduces cell growth. Our integrative approach thus links glutamine addiction to glutamate excretion in cancer and points toward potential drug targets.

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“…This study investigated the effects of two concentrations of L-cystine representing physiological condition and routine culture medium on ferroptosis induced by sulfasalazine, erastin, and RSL3. Ferroptosis was determined by cell viability, membrane permeabilization, and lipid peroxidation in Huh6, Huh7, and SK-HEP-1 hepatocellular carcinoma cell lines which were susceptible to ferroptotic cell death in our previous study ( Kim et al ., 2019 ; Yuk et al ., 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Extracellular Concentration of _L-Cystine Determines the Sensitivity to System x_c^- Inhibitors

Abdullah

Lee

2021

Biomol Ther (Seoul)

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Targeting the cystine/glutamate exchange transporter, system xc -, is a promising anticancer strategy that induces ferroptosis, which is a distinct form of cell death mediated by iron-dependent lipid peroxidation. The concentration of L-cystine in culture medium is higher than the physiological level. This study was aimed to evaluate the effects of L-cystine concentration on the efficacy of ferroptosis inducers in hepatocellular carcinoma cells. This study showed that treatment with sulfasalazine or erastin, a system xcinhibitor, decreased the viability of Huh6 and Huh7 cells in a dose-dependent manner, and the degree of growth inhibition was greater in medium containing a physiological L-cystine concentration of 83 µM than in commercial medium with a concentration of 200 µM L-cystine. However, RSL3, a glutathione peroxidase 4 inhibitor, decreased cell viability to a similar extent in media containing both L-cystine concentrations. Sulfasalazine and erastin significantly increased the percentages of propidium iodide-positive cells in media with 83 µM L-cystine, but not in media with 200 µM L-cystine. Sulfasalazine-or erastin-induced accumulation of lipid peroxidation as monitored by C11-BODIPY probe was higher in media with 83 µM L-cystine than in media with 200 µM L-cystine. In contrast, the changes in the percentages of propidium iodide-positive cells and lipid peroxidation by RSL3 were similar in both media. These results showed that sulfasalazine and erastin, but not RSL3, were efficacious under conditions of physiological Lcystine concentration, suggesting that medium conditions would be crucial for the design of a bioassay for system xcinhibitors.

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Mechanisms of Growth Inhibition by Sulfasalazine and Erastin in Hepatocellular Carcinoma Cell Lines

Cited by 3 publications

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Necrostatin-1 Prevents Ferroptosis in a RIPK1- and IDO-Independent Manner in Hepatocellular Carcinoma

Necrostatin-1 Prevents Ferroptosis in a RIPK1- and IDO-Independent Manner in Hepatocellular Carcinoma

Quantitative analysis of amino acid metabolism in liver cancer links glutamate excretion to nucleotide synthesis

Extracellular Concentration of _L-Cystine Determines the Sensitivity to System x_c^- Inhibitors

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Mechanisms of Growth Inhibition by Sulfasalazine and Erastin in Hepatocellular Carcinoma Cell Lines

Cited by 3 publications

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Necrostatin-1 Prevents Ferroptosis in a RIPK1- and IDO-Independent Manner in Hepatocellular Carcinoma

Necrostatin-1 Prevents Ferroptosis in a RIPK1- and IDO-Independent Manner in Hepatocellular Carcinoma

Quantitative analysis of amino acid metabolism in liver cancer links glutamate excretion to nucleotide synthesis

Extracellular Concentration of L-Cystine Determines the Sensitivity to System xc- Inhibitors

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Extracellular Concentration of _L-Cystine Determines the Sensitivity to System x_c^- Inhibitors