Journal: Redox Biology

Article Title: HDAC inhibition induces EMT and alterations in cellular iron homeostasis to augment ferroptosis sensitivity in SW13 cells

doi: 10.1016/j.redox.2021.102149

Figure Lengend Snippet: HDAC inhibitor mediated EMT in SW13 cells is not influenced by changes in cellular iron availability. (A) EMT-like changes in actin (green) morphology induced by treatment with 2 nM FK228 are not altered by iron chelation (FK228 + DFO) or iron supplementation (FK228 + Hemin). (B) Increased expression of the mesenchymal marker, vimentin by FK228 treatment is not influenced by co-treatment with DFO or hemin. (C) Increased mRNA expression of the mesenchymal markers SMARCA2, TGFβ1, and SNAI1 following HDAC inhibitor treatment were not influence by iron chelation. (D) The mRNA expression of matrix metalloproteinase enzymes MMP2 and MMP9 was also increased by HDAC inhibitor treatment and unaffected by iron chelation. (E) Increased intracellular iron accumulation following HDAC inhibitor mediated EMT (FK228) were consistent with levels observed following iron supplementation (Hemin). Images were taken using a 40X objective lens. Data are presented as mean ± SEM. * Denotes significant difference compared to control, p < 0.05. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: For EMT induction, SW13 cells were treated with 2 nM Romidepsin (FK228, Selleckchem, Houston, TX, USA) for 48 h. To examine the influence of iron availability on EMT progression, SW13 cells were treated with 2 nM FK228 for 24 h and then co-treated with 50 μM desferrioxamine (DFO), an iron chelator, or 40 μM hemin, an iron-containing porphyrin, for another 24 h. To induce ferroptosis, cells were treated with either 1 μM or 5 μM erastin for 24 or 48 h. All experiments followed the described treatment dose and length unless mentioned otherwise in the methods session.

Techniques: Expressing, Marker, Control

Journal: Redox Biology

Article Title: HDAC inhibition induces EMT and alterations in cellular iron homeostasis to augment ferroptosis sensitivity in SW13 cells

doi: 10.1016/j.redox.2021.102149

Figure Lengend Snippet: IRP mRNA binding activity is increased and ferroportin expression is decreased following HDAC inhibitor mediated EMT in SW13 cells. Spontaneous (A) and total (B) mRNA binding activity and expression of the IRP target proteins TFRC and ferroportin (C) in SW13 cells that were treated with DMSO (Control) or with 2 nM FK228 for 24 h were measured by gel-shift assay and Western blot, respectively. GAPDH was used as a loading control. Data are presented as mean ± SEM. * Denotes significant difference compared to control, p < 0.05.

Article Snippet: For EMT induction, SW13 cells were treated with 2 nM Romidepsin (FK228, Selleckchem, Houston, TX, USA) for 48 h. To examine the influence of iron availability on EMT progression, SW13 cells were treated with 2 nM FK228 for 24 h and then co-treated with 50 μM desferrioxamine (DFO), an iron chelator, or 40 μM hemin, an iron-containing porphyrin, for another 24 h. To induce ferroptosis, cells were treated with either 1 μM or 5 μM erastin for 24 or 48 h. All experiments followed the described treatment dose and length unless mentioned otherwise in the methods session.

Techniques: Binding Assay, Activity Assay, Expressing, Control, Gel Shift, Western Blot

Journal: Redox Biology

Article Title: HDAC inhibition induces EMT and alterations in cellular iron homeostasis to augment ferroptosis sensitivity in SW13 cells

doi: 10.1016/j.redox.2021.102149

Figure Lengend Snippet: Both iron chelation and iron supplementation reduce cell viability in HDAC inhibitor treated SW13 cells. (A) Images and (B) quantitation of EdU-positive cells in SW13 cells following treatment with DMSO (Control) or 2 nM FK228 treatment for 24 h and then co-treatment with 50 μM DFO (FK228 + DFO) or 40 μM hemin (FK228 + Hemin) for another 24 h. (C) Percent cell viability was measured by fluorometric assay. Images were taken through an 4X objective lens. Data are presented as mean ± SEM. *Denotes statistical significance, p < 0.05.

Article Snippet: For EMT induction, SW13 cells were treated with 2 nM Romidepsin (FK228, Selleckchem, Houston, TX, USA) for 48 h. To examine the influence of iron availability on EMT progression, SW13 cells were treated with 2 nM FK228 for 24 h and then co-treated with 50 μM desferrioxamine (DFO), an iron chelator, or 40 μM hemin, an iron-containing porphyrin, for another 24 h. To induce ferroptosis, cells were treated with either 1 μM or 5 μM erastin for 24 or 48 h. All experiments followed the described treatment dose and length unless mentioned otherwise in the methods session.

Techniques: Quantitation Assay, Control

Journal: Redox Biology

Article Title: HDAC inhibition induces EMT and alterations in cellular iron homeostasis to augment ferroptosis sensitivity in SW13 cells

doi: 10.1016/j.redox.2021.102149

Figure Lengend Snippet: HDAC inhibitor-converted mesenchymal-like SW13 cells have increased levels of ROS and reduced expression of antioxidant defense genes. (A) ROS levels in SW13 following treatment with DMSO (Control) or 2 nM FK228 treatment were measured by incubating cells with 5 μM CellRox deep red reagent for 30 min and (B) quantitated relative to Hoechst stained nuclei with ImageJ. Images were taken through an 40X objective lens. (C) The relative mRNA abundance of SOD2, SLC7A11, TP53, and GPX4 were significantly decreased by FK228 treatment. Data are presented as mean ± SEM. *Denotes significance compared to control, p < 0.05. (For interpretation of the references to color in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: For EMT induction, SW13 cells were treated with 2 nM Romidepsin (FK228, Selleckchem, Houston, TX, USA) for 48 h. To examine the influence of iron availability on EMT progression, SW13 cells were treated with 2 nM FK228 for 24 h and then co-treated with 50 μM desferrioxamine (DFO), an iron chelator, or 40 μM hemin, an iron-containing porphyrin, for another 24 h. To induce ferroptosis, cells were treated with either 1 μM or 5 μM erastin for 24 or 48 h. All experiments followed the described treatment dose and length unless mentioned otherwise in the methods session.

Techniques: Expressing, Control, Staining

Journal: Redox Biology

Article Title: HDAC inhibition induces EMT and alterations in cellular iron homeostasis to augment ferroptosis sensitivity in SW13 cells

doi: 10.1016/j.redox.2021.102149

Figure Lengend Snippet: HDAC inhibitor treatment promotes anti-apoptotic mRNA expression in SW13 cells. The relative abundance of (A) ferroptotic, (B) pro-apoptotic, and (C) anti-apoptotic mRNAs were measured by qPCR following treatment with DMSO (Control), 2 nM FK228, 5 μM erastin, or a combination of 2 nM FK228 and 1 μM erastin for 48 h. Data are presented as mean ± SEM. *Denotes significance compared to control, p < 0.05.

Article Snippet: For EMT induction, SW13 cells were treated with 2 nM Romidepsin (FK228, Selleckchem, Houston, TX, USA) for 48 h. To examine the influence of iron availability on EMT progression, SW13 cells were treated with 2 nM FK228 for 24 h and then co-treated with 50 μM desferrioxamine (DFO), an iron chelator, or 40 μM hemin, an iron-containing porphyrin, for another 24 h. To induce ferroptosis, cells were treated with either 1 μM or 5 μM erastin for 24 or 48 h. All experiments followed the described treatment dose and length unless mentioned otherwise in the methods session.

Techniques: Expressing, Control

Journal: NPJ Precision Oncology

Article Title: Multi-modal characterization of metabolic and immune gene clusters in adrenocortical carcinoma treatment

doi: 10.1038/s41698-025-01092-4

Figure Lengend Snippet: a, b KEGG and HALLMARK GSEA in SIS subgroups. c Common genes screened from the core genes of the two enrichment pathways. d The distribution of the ensemble of ADS, hormone, and cortisol levels among SIS subgroups. e Correlation between SIS and ADS. f, g SIS and ADS are distributed over hormones and cortisol, respectively. h Correlation of steroid hormone-related genes and Mitotane primary target genes with SIS and ADS expression. Expression values were compared with low SIS, high ADS, hormone present, and cortisol present. i Lasso regression, RF, and SVM-RFE machine learning jointly screened for genes most associated with SIS. j Kaplan-Meier curve was used to predict DHCR7 overall survival. k Protein expression of DHCR7 in ACC tissues and normal adrenal tissues by IHC. l Distribution of DHCR7 expression in single-cell RNA sequencing. m Mechanistic pathways associated with DHCR7 and Mitotane. n Cell viability was assessed after treatment with different concentrations of Mitotane in SW-13 cells. Control or DHCR7 siRNA was transfected, incubated for 48 h, and then collected for RT-PCR analysis of the DHCR7 gene. Cell viability was assessed after treatment of SW-13 cells with control or DHCR7 siRNA transfected with Mitotane (6.9 μM) for 48 h. o Cell viability was evaluated in NCI-H295R cells treated with varying concentrations of Mitotane, as well as in cells transfected with control or DHCR7 siRNA and incubated for 48 h, followed by RT-PCR analysis of DHCR7 expression. Cell viability was assessed after treatment of NCI-H295R cells with control, or DHCR7 siRNA transfected with Mitotane (4.7 μM) for 48 hours. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Procell Life Science & Technology provided human ACC cell lines (SW13, NCI-H296R).

Techniques: Expressing, RNA Sequencing, Control, Transfection, Incubation, Reverse Transcription Polymerase Chain Reaction