Journal: Cell Communication and Signaling : CCS

Article Title: GPX3 promotes cisplatin resistance in TNBC by manipulating ROS-TGFB1-ZEB2

doi: 10.1186/s12964-025-02356-z

Figure Lengend Snippet: GPX3 enhanced TGF-β1 signaling following cisplatin treatment in MDA-MB-231 cells. (A-B) RT-qPCR analysis of TGFB1 signaling and its EMT marker expression levels in cells transfected with NC or GPX3-KD, treated with or without rhTGFβ1. (C) Western blot analysis of TGFB1 signaling and EMT marker expression levels in cells transfected with NC or GPX3-KD, treated with or without rhTGFβ1. (D-E) Immunofluorescence staining for SMAD2 and SMAD3 in the nuclear and cytoplasmic compartments of NC and GPX3-KD cells treated with rhTGFβ1, with corresponding quantification. (F) Western blot analysis of the ratio of phosphorylated SMAD2/3 (p-SMAD2/3) to total SMAD2 and SMAD3 in NC and GPX3-KD cells treated with rhTGFβ1. (G-H) Secretion levels of total and active TGFB1 in the supernatants of GPX3-KD cells incubated with cisplatin, with or without NAC pre-treatment, as detected by Western blot and ELISA. (I) Western blot analysis of TGF-β signaling and EMT marker expression in GPX3-KD cells, with or without NAC pre-treatment. (ns P ≥ 0.05, *P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: Cisplatin (Cat. No. HY-17394), SB431542 (TGF-β receptor kinase inhibitors, TRKI, HY-10431), and recombinant human TGFβ1 protein from HEK293 (Cat. No. HY- P70543 ) were purchased from MedChemExpress (MCE).

Techniques: Quantitative RT-PCR, Marker, Expressing, Transfection, Western Blot, Immunofluorescence, Staining, Incubation, Enzyme-linked Immunosorbent Assay

Journal: Cell Communication and Signaling : CCS

Article Title: GPX3 promotes cisplatin resistance in TNBC by manipulating ROS-TGFB1-ZEB2

doi: 10.1186/s12964-025-02356-z

Figure Lengend Snippet: GPX3-TGFB1 promoted ZEB2 expression and enhanced EMT during cisplatin treatment in TNBC. (A) GEPIA analysis of the relationship between the expression of GPX3, TGFB1, and ZEB2. (B) Correlation analysis of the protein expression levels of GPX3, TGFB1, and ZEB2 in TNBC patient tissue microarrays (n=75). (C-D) Western blot analysis showed that TGFB1 reversed the effect of GPX3 on ZEB2 expression. (E) Existing ChIP-seq analysis of SMAD3 binding sites on the ZEB2 gene promoter (ChiP-Atlas: Enrichment Analysis, https://chip-atlas.org/enrichment_analysis). (F) ChIP-qPCR validation of SMAD3 enrichment at the ZEB2 gene promoter. (G-I) Knocking down SMAD2 or SMAD3 or inhibiting TGFBR1 kinase activity abolished rhTGFβ1-induced transcriptional activity (G), expression level of ZEB2 (H), and EMT markers (I). (ns P ≥ 0.05, *P < 0.05, **P < 0.01, ***P < 0.001)

Article Snippet: Cisplatin (Cat. No. HY-17394), SB431542 (TGF-β receptor kinase inhibitors, TRKI, HY-10431), and recombinant human TGFβ1 protein from HEK293 (Cat. No. HY- P70543 ) were purchased from MedChemExpress (MCE).

Techniques: Expressing, Western Blot, ChIP-sequencing, Binding Assay, ChIP-qPCR, Biomarker Discovery, Activity Assay