Journal: Cell Death & Disease

Article Title: Selenomethionine as a dual-mechanism ferroptosis inhibitor: selenium-supply-driven GPX4 biosynthesis beyond transsulfuration and reductive-capacity-mediated ROS scavenging independent of GPX4 activity

doi: 10.1038/s41419-026-08466-x

Figure Lengend Snippet: a Schematic of the small molecule screening strategy. b The small molecule screening results were exhibited as the reduced ratio of PI and the recovery ratio of Hoechst in small molecule-treated cells versus vehicle-treated cells. After cell seeding, the HT1080 cells were incubated with the FDA drug (2625) or DMSO and 0.2 μM RSL3 for 24 h. Then, the cell was measured with PI and Hoechst using CX7 LED Pro. c The small molecule screening results were exhibited as the reduced ratio of PI and the recovery ratio of Hoechst in small molecule-treated cells versus vehicle-treated cells. After cell seeding, the HT1080 cells were incubated with the natural product (3651) or DMSO and 0.2 μM RSL3 for 24 h. Then, the cell was measured with PI and Hoechst using CX7 LED Pro. d , e HT1080 and OS-RC-2 cells were treated with RSL3 (2 μM) or selenomethionine (SeleMet) (100 μM) as indicated for 4–6 h ( n = 3). d The lipid ROS levels were determined using flow cytometry (HT1080 and OS-RC-2 for 4 h) ( n = 3). e The cell death levels were quantified using flow cytometry (HT1080 and OS-RC-2 for 6 h) ( n = 3). f The cell viability of HT1080 cells that were treated with RSL3 and SeleMet (100 μM) for 6 h ( n = 3). g , h HT1080 cells are cultured in ± CC (cystine) medium treated with SeleMet (100 μM) or Ferrostatin-1 (Fer-1, 10 μM) as indicated for 10-14 h ( n = 3). g The lipid ROS levels were determined using flow cytometry (HT1080 for 10 h) ( n = 3). h The cell death levels were quantified using flow cytometry (HT1080 for 14 h) ( n = 3). CC, cystine, 0.4 mM; Fer-1, Ferrostatin-1, 10 μM. Data were represented as mean ± SD with P values determined by one-way ANOVA (d, e, g, and h). *** P < 0.001; **** P < 0.0001; ns, nonsignificant.

Article Snippet: The compounds used are Ferrostain-1 (Fer-1) (S7243, Selleck), Propidium iodide (PI) (P4170, Sigma), BODIPY581/591 C11 (D3861, ThermoFisher), DCFH2-DA (D6883, Sigma), Cystine (30200, Sigma), Methionine (Met) (M9625, Sigma), RhoNox-1 (HY-D1533, MCE), α-Vitamin E ((+)-α-Tocopherol) (HY-N0683, MCE), RSL3 (HY-100218A, MCE), DL-Propargylglycine (HY-W040124, MCE), Selenomethionine (HY-B1000, MCE), L-Selenocystine (HY-129960, MCE), Cisplatin (HY-17394, MCE), Cycloheximide (HY-12320, MCE), FDA-Approved Drug Library (HY-L022, MCE), Natural Product Library (HY-L021, MCE), Dulbecco’s Modified Eagle’s Medium high glucose without L-methionine, L-cystine, and L-glutamine (D0422-100 ml, Sigma), and Penicillin-Streptomycin (P06-07100, Pan Biotech).

Techniques: Incubation, Flow Cytometry, Cell Culture

Journal: Cell Death & Disease

Article Title: Selenomethionine as a dual-mechanism ferroptosis inhibitor: selenium-supply-driven GPX4 biosynthesis beyond transsulfuration and reductive-capacity-mediated ROS scavenging independent of GPX4 activity

doi: 10.1038/s41419-026-08466-x

Figure Lengend Snippet: a Schematic of metabolic processes of selenium containing amino acids. b The protein levels of the GPX4 in OS-RC-2, HepG2, and HT1080 cells are cultured in ± CC (cystine) medium treated with SeleMet were measured by western blot (WB). c The protein levels of the GPX4 in OS-RC-2, HepG2, and HT1080 cells are cultured in medium treated with RSL3 or SeleMet were measured by WB. d The protein levels of the GPX4 in HepG2 and HT1080 cells are cultured in medium treated with SeleMet or DL-Propargylglycine (PAG, 100 μM) were measured by WB. e The protein levels of the GPX4 in HepG2 cells are cultured in ± CC (cystine) medium treated with SeleMet or PAG were measured by WB. f , g HT1080 cells are cultured in ± CC (cystine) medium treated with SeleMet, PAG, or fer-1 as indicated for 10-14 h ( n = 3). f The lipid ROS levels were determined using flow cytometry (HT1080 for 10 h) ( n = 3). g The cell death levels were quantified using flow cytometry (HT1080 for 14 h) ( n = 3). h The protein levels of the CBS and CTH in HepG2 cells treated with shCBS and shCTH were measured by WB. i , j HepG2 (shNC, shCBS, and shCTH) cells are treated with RSL3 or SeleMet as indicated for 4-6 h ( n = 3). i The lipid ROS levels were determined using flow cytometry (for 4 h) ( n = 3). j The cell death levels were quantified using flow cytometry (for 6 h) ( n = 3). k Analysis of the transformation from selenocystine to selenocysteine using DNTB. The transformation reaction is catalyzed by GSR and utilizes NADPH as the reducing force. l HT1080 and OS-RC-2 cells are cultured in ± CC (cystine) medium treated with selenomethionine or selenocystine as indicated for 10–24 h, then the total SH+SeH is measured by DNTB ( n = 3). CC, cystine, 0.4 mM; Fer-1, Ferrostatin-1, 10 μM; PAG, DL-Propargylglycine. Data were represented as mean ± SD with P values determined by one-way ANOVA (f, g, i, j, k, and l). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, nonsignificant.

Article Snippet: The compounds used are Ferrostain-1 (Fer-1) (S7243, Selleck), Propidium iodide (PI) (P4170, Sigma), BODIPY581/591 C11 (D3861, ThermoFisher), DCFH2-DA (D6883, Sigma), Cystine (30200, Sigma), Methionine (Met) (M9625, Sigma), RhoNox-1 (HY-D1533, MCE), α-Vitamin E ((+)-α-Tocopherol) (HY-N0683, MCE), RSL3 (HY-100218A, MCE), DL-Propargylglycine (HY-W040124, MCE), Selenomethionine (HY-B1000, MCE), L-Selenocystine (HY-129960, MCE), Cisplatin (HY-17394, MCE), Cycloheximide (HY-12320, MCE), FDA-Approved Drug Library (HY-L022, MCE), Natural Product Library (HY-L021, MCE), Dulbecco’s Modified Eagle’s Medium high glucose without L-methionine, L-cystine, and L-glutamine (D0422-100 ml, Sigma), and Penicillin-Streptomycin (P06-07100, Pan Biotech).

Techniques: Cell Culture, Western Blot, Flow Cytometry, Transformation Assay

Journal: Cell Death & Disease

Article Title: Selenomethionine as a dual-mechanism ferroptosis inhibitor: selenium-supply-driven GPX4 biosynthesis beyond transsulfuration and reductive-capacity-mediated ROS scavenging independent of GPX4 activity

doi: 10.1038/s41419-026-08466-x

Figure Lengend Snippet: a Schematic of metabolic processes of selenomethionine (SeleMet). b The mRNA levels of the indicated metabolic enzymes in HT1080 were in ± cystine (Cys 2 ) medium treated with SeleMet and shown in heatmap. Red color indicated high expression, and blue indicated low expression. c , d HT1080 cells are cultured medium treated with RSL3, SeleMet, FIDAS-5, PAG, or fer-1 as indicated for 4-6 h ( n = 3). c The lipid ROS levels were determined using flow cytometry (HT1080 for 4 h) ( n = 3). d The cell death levels were quantified using flow cytometry (HT1080 for 6 h) ( n = 3). e The cell viability of HT1080 cells that were treated with RSL3, SeleMet, FIDAS-5, PAG, or fer-1 for 6 h ( n = 3). f The protein levels of the GPX4 in 293 T and HT1080 treated with sgGPX4 were measured by WB. g , h 293 T and HT1080 KO GPX4 cells are cultured medium treated with Vitamin E (VitE) or SeleMet as indicated for 10–14 h ( n = 3). g The lipid ROS levels were determined using flow cytometry (for 10 h) ( n = 3). h The cell death levels were quantified using flow cytometry (for 14 h) ( n = 3). Data were represented as mean ± SD with P values determined by one-way ANOVA ( c , d , g , and h ). * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001; ns, nonsignificant.

Article Snippet: The compounds used are Ferrostain-1 (Fer-1) (S7243, Selleck), Propidium iodide (PI) (P4170, Sigma), BODIPY581/591 C11 (D3861, ThermoFisher), DCFH2-DA (D6883, Sigma), Cystine (30200, Sigma), Methionine (Met) (M9625, Sigma), RhoNox-1 (HY-D1533, MCE), α-Vitamin E ((+)-α-Tocopherol) (HY-N0683, MCE), RSL3 (HY-100218A, MCE), DL-Propargylglycine (HY-W040124, MCE), Selenomethionine (HY-B1000, MCE), L-Selenocystine (HY-129960, MCE), Cisplatin (HY-17394, MCE), Cycloheximide (HY-12320, MCE), FDA-Approved Drug Library (HY-L022, MCE), Natural Product Library (HY-L021, MCE), Dulbecco’s Modified Eagle’s Medium high glucose without L-methionine, L-cystine, and L-glutamine (D0422-100 ml, Sigma), and Penicillin-Streptomycin (P06-07100, Pan Biotech).

Techniques: Expressing, Cell Culture, Flow Cytometry

Journal: Cell Death & Disease

Article Title: Selenomethionine as a dual-mechanism ferroptosis inhibitor: selenium-supply-driven GPX4 biosynthesis beyond transsulfuration and reductive-capacity-mediated ROS scavenging independent of GPX4 activity

doi: 10.1038/s41419-026-08466-x

Figure Lengend Snippet: a The structural formula of methionine (Met) and selenomethionine (SeleMet). b , c HT1080 cells are cultured ± cystine (CC) medium treated with met, seleMet, or fer-1 as indicated for 10-14 h ( n = 3). b The lipid ROS levels were determined using flow cytometry (HT1080 for 10 h) ( n = 3). c The cell death levels were quantified using flow cytometry (HT1080 for 14 h) ( n = 3). d HT1080 cells are cultured ± cystine (CC) medium treated with met, seleMet, or fer-1 as indicated for 10 h, then the ROS levels were determined using flow cytometry ( n = 3). e Analysis of the reducibility of GSH and seleMet using 2,2-Diphenyl-1-picrylhydrazyl (DPPH), which has a single electron and can accept one electron or hydrogen ion, with maximum absorption at a wavelength of 517 nm ( n = 3). f The flowchart of chemical reaction and identification. g Mass spectrometry detection results of selenometionine and selenomethionine sulfoxide. Data were represented as mean ± SD with P values determined by one-way ANOVA ( b – d ). *** P < 0.001; **** P < 0.0001; ns, nonsignificant.

Article Snippet: The compounds used are Ferrostain-1 (Fer-1) (S7243, Selleck), Propidium iodide (PI) (P4170, Sigma), BODIPY581/591 C11 (D3861, ThermoFisher), DCFH2-DA (D6883, Sigma), Cystine (30200, Sigma), Methionine (Met) (M9625, Sigma), RhoNox-1 (HY-D1533, MCE), α-Vitamin E ((+)-α-Tocopherol) (HY-N0683, MCE), RSL3 (HY-100218A, MCE), DL-Propargylglycine (HY-W040124, MCE), Selenomethionine (HY-B1000, MCE), L-Selenocystine (HY-129960, MCE), Cisplatin (HY-17394, MCE), Cycloheximide (HY-12320, MCE), FDA-Approved Drug Library (HY-L022, MCE), Natural Product Library (HY-L021, MCE), Dulbecco’s Modified Eagle’s Medium high glucose without L-methionine, L-cystine, and L-glutamine (D0422-100 ml, Sigma), and Penicillin-Streptomycin (P06-07100, Pan Biotech).

Techniques: Cell Culture, Flow Cytometry, Mass Spectrometry

Journal: Cell Death & Disease

Article Title: Selenomethionine as a dual-mechanism ferroptosis inhibitor: selenium-supply-driven GPX4 biosynthesis beyond transsulfuration and reductive-capacity-mediated ROS scavenging independent of GPX4 activity

doi: 10.1038/s41419-026-08466-x

Figure Lengend Snippet: a The mice were injected with cisplatin (20 mg/kg) only or combined with selenomethionine (3 mg/kg) or Fer-1 (1 mg/kg) as indicated for 4 days. b Morphology of mice after drug treatment. c Morphology of kidneys after drug treatment. d Kidney/body weight ratio after drug treatment. e The blood urea nitrogen (BUN) of mice after drug treatment. f The serum creatinine of mice after drug treatment. g The MDA of kidney after drug treatment. h WB detection of GPX4 protein levels in mice kidneys after drug treatment. i HE staining of kidney after drug treatment. j PAS staining of kidney after drug treatment. k F4/80 staining of kidney after drug treatment. Data were represented as mean ± SD with P values determined by one-way ANOVA ( d – g ). **** P < 0.0001; ns, nonsignificant.

Article Snippet: The compounds used are Ferrostain-1 (Fer-1) (S7243, Selleck), Propidium iodide (PI) (P4170, Sigma), BODIPY581/591 C11 (D3861, ThermoFisher), DCFH2-DA (D6883, Sigma), Cystine (30200, Sigma), Methionine (Met) (M9625, Sigma), RhoNox-1 (HY-D1533, MCE), α-Vitamin E ((+)-α-Tocopherol) (HY-N0683, MCE), RSL3 (HY-100218A, MCE), DL-Propargylglycine (HY-W040124, MCE), Selenomethionine (HY-B1000, MCE), L-Selenocystine (HY-129960, MCE), Cisplatin (HY-17394, MCE), Cycloheximide (HY-12320, MCE), FDA-Approved Drug Library (HY-L022, MCE), Natural Product Library (HY-L021, MCE), Dulbecco’s Modified Eagle’s Medium high glucose without L-methionine, L-cystine, and L-glutamine (D0422-100 ml, Sigma), and Penicillin-Streptomycin (P06-07100, Pan Biotech).

Techniques: Injection, Staining