apoptosis inhibitor 20  (MedChemExpress)

Journal: Cancers

Article Title: NSUN4 Suppresses Ferroptosis Through m 5 C-Dependent Stabilization of C-MYC and Activation of the PI3K/Akt Signaling Pathway in Cervical Cancer

doi: 10.3390/cancers18091392

Figure Lengend Snippet: NSUN4 is upregulated in cervical cancer and associated with poor patient prognosis. ( A ) Differential expression analysis of TCGA RNA-sequencing data showing that NSUN4 is significantly upregulated in cervical cancer tissues compared with normal tissues. ( B ) Kaplan–Meier survival curves demonstrating that patients with high NSUN4 expression exhibit significantly worse progression-free survival than those with low expression ( p < 0.05). ( C ) Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses indicating that NSUN4-associated genes are mainly involved in apoptosis, PI3K–Akt signaling, and oxidative phosphorylation pathways. ( D , E ) Validation of NSUN4 expression in paired cervical cancer and adjacent normal tissues by qRT-PCR and Western blot, confirming elevated NSUN4 levels at both mRNA and protein levels in tumor samples . The arrow indicates the specific NSUN4 band. ( F ) Representative NSUN4 IHC staining images of cervical cancer and matched adjacent normal tissues from a tissue microarray (TMA). ( G ) Analysis of IHC scores from a cohort of 30 cervical cancer patients, showing the comparison of NSUN4 expression between cervical cancer and matched adjacent normal tissues. Statistical analysis was performed using a paired Student’s t -test (*** p < 0.001).

Article Snippet: To assess the involvement of ferroptosis and the PI3K/AKT signaling pathway, cells were treated for 24 h with the apoptosis inhibitor Z-VAD-FMK (MedChemExpress, Monmouth Junction, NJ, USA), the necroptosis inhibitor Nec-1 (MedChemExpress, Monmouth Junction, NJ, USA), the ferroptosis inhibitors Ferrostatin-1 (MedChemExpress, Monmouth Junction, NJ, USA) and Liproxstatin-1 (MedChemExpress, Monmouth Junction, NJ, USA), or 3-Methyladenine (MedChemExpress, Monmouth Junction, NJ, USA), which was used in this study as an inhibitor of the PI3K pathway.

Techniques: Quantitative Proteomics, RNA Sequencing, Expressing, Phospho-proteomics, Biomarker Discovery, Quantitative RT-PCR, Western Blot, Immunohistochemistry, Microarray, Comparison

Journal: Cancers

Article Title: NSUN4 Suppresses Ferroptosis Through m 5 C-Dependent Stabilization of C-MYC and Activation of the PI3K/Akt Signaling Pathway in Cervical Cancer

doi: 10.3390/cancers18091392

Figure Lengend Snippet: NSUN4 promotes cervical cancer cell proliferation by suppressing ferroptosis. ( A ) KEGG pathway enrichment analysis of transcriptome sequencing data from NSUN4-overexpressing and control cells revealing a significant association with ferroptosis pathways. ( B ) Cell viability assays showing that ferroptosis inhibitors (Fer-1 and Lipro-1), but not apoptosis or necroptosis inhibitors (Z-VAD-FMK or Nec-1), rescue the proliferation defects induced by NSUN4 knockdown. ( C , D ) Western blot analysis demonstrating that NSUN4 depletion downregulates, whereas NSUN4 overexpression upregulates, ferroptosis-associated proteins SLC7A11, FTH1, and GPX4. ( E – G ) Quantification of intracellular GSH levels showing that NSUN4 overexpression increases, while NSUN4 knockdown decreases, GSH content; ferroptosis inhibition with Fer-1 or Lipro-1 restores GSH levels in NSUN4-deficient cells. Data are presented as mean ± SD. ns, not significant; * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: To assess the involvement of ferroptosis and the PI3K/AKT signaling pathway, cells were treated for 24 h with the apoptosis inhibitor Z-VAD-FMK (MedChemExpress, Monmouth Junction, NJ, USA), the necroptosis inhibitor Nec-1 (MedChemExpress, Monmouth Junction, NJ, USA), the ferroptosis inhibitors Ferrostatin-1 (MedChemExpress, Monmouth Junction, NJ, USA) and Liproxstatin-1 (MedChemExpress, Monmouth Junction, NJ, USA), or 3-Methyladenine (MedChemExpress, Monmouth Junction, NJ, USA), which was used in this study as an inhibitor of the PI3K pathway.

Techniques: Sequencing, Control, Knockdown, Western Blot, Over Expression, Inhibition