Journal: Molecular Therapy. Nucleic Acids

Article Title: Combining anti-gene γPNA with small molecules and RNA inhibitors: A strategy to enhance anti-tumor efficacy

doi: 10.1016/j.omtn.2025.102804

Figure Lengend Snippet: Anti-transcription activity of γPNA1 with HDACi in lymphoma cells Relative fold change of c-Myc levels in U2932 cells measured by real-time PCR on day 2 after treatment with (A) γPNA1 and (B) ScR-γPNA2 in combination with romidepsin, entinostat, vorinostat, panobinostat, and belinostat. Results are presented as mean ± SEM and two-way ANOVA was used to determine the statistically significant difference between groups. Western blot analysis representing the change in c-MYC protein on day 2 after treatment with γPNA1 and ScR-γPNA2 in combination with (C) romidepsin, (D) entinostat, (E) vorinostat, (F) panobinostat, and (G) belinostat. ∗∗(C–F) Cyclophilin B was used as an endogenous control, and the same blots are presented in C–S3G. c-MYC, EZH2, and cyclophilin B were probed from the same blot. Results are presented as mean ± SEM, and the p value between groups was determined using one-way ANOVA.

Article Snippet: Taqman gene expression assay for c-Myc (Thermo Fisher Scientific, Hs00153408), GAPDH (Thermo Fisher Scientific, Hs02758991), and EZH2 (Thermo Fisher Scientific, Hs00544830) was used to amplify the respective mRNAs using the specified cycling conditions in the QuantStudio 5 real-time PCR detection system (Thermo Fisher Scientific, USA).

Techniques: Activity Assay, Real-time Polymerase Chain Reaction, Western Blot, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: Combining anti-gene γPNA with small molecules and RNA inhibitors: A strategy to enhance anti-tumor efficacy

doi: 10.1016/j.omtn.2025.102804

Figure Lengend Snippet: MYC/MAX inhibitors in combination with anti-transcription γPNA1 Cell viability of (A) U2932 and (B) Raji cells treated with increasing doses of MYC/MAX inhibitors (Myci975, EN4, 10058-F4, and sAJM589) alone and in combination with γPNA1 and ScR-γPNA2 (8 μM) for 72 h. Results are presented as mean ± SEM. The IC 50 (95% CI) values of MYC/MAX inhibitors alone and combination treatment of MYC/MAX with γPNA1 in (C) U2932 and (D) Raji cells. (E) Cell viability of γPNA1-treated U2932 and Raji cells at 8 μM concentration. Western blot analysis representing the change in c-MYC protein 72 h after treatment with γPNA1 and ScR-γPNA2 in combination with (F) Myci975, (G) EN4, (H) 10058-F4, and (I) sAJM589. ∗∗(F–I) Cyclophilin B was used as an endogenous control, and the same blots are presented in A–S7D. c-MYC, EZH2, and cyclophilin B were probed from the same blot. Results are presented as mean ± SEM, p value for one-way ANOVA.

Article Snippet: Taqman gene expression assay for c-Myc (Thermo Fisher Scientific, Hs00153408), GAPDH (Thermo Fisher Scientific, Hs02758991), and EZH2 (Thermo Fisher Scientific, Hs00544830) was used to amplify the respective mRNAs using the specified cycling conditions in the QuantStudio 5 real-time PCR detection system (Thermo Fisher Scientific, USA).

Techniques: Concentration Assay, Western Blot, Control

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: Elevated EZH2 expression correlates with poor prognosis in hepatocellular carcinoma (HCC). A Expression of EZH2 in the TCGA-HCC dataset, comparing HCC tissues with peri-tumoral liver tissues. B RT-qPCR assays demonstrating mRNA levels of EZH2 in 12 HCC patient samples and their corresponding adjacent normal tissues. C , D Western blot demonstrating protein levels of EZH2 in 12 HCC patient samples and their corresponding adjacent normal tissues. E Representative immunohistochemistry (IHC) images showing EZH2 expression in HCC tissues versus paired adjacent normal tissues. Statistical analysis comparing IHC scores of 12 HCC tissues with their paired normal tissues. F , G Kaplan-Meier overall survival curve and disease-free survival curve of HCC patients correlated with the expression of EZH2. All data are presented as the means ± SDs, n = 5 per group, * ( p < 0.05), ** ( p < 0.01), ns = no significance

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Immunohistochemistry

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: EZH2 expression is increased in lenvatinib-resistant HCC cells and contributes to resistance development. A Schematic representation of the establishment of lenvatinib-resistant (LR) cells. B Cell viability assessed by CCK-8 assay following exposure to varying concentrations of lenvatinib for 48 h. Proliferation of both Huh7-LR and Hep3B-LR cells was greater than that of their parental cells at different lenvatinib concentrations. C , D mRNA and protein levels of EZH2 in Huh7-LR and Hep3B-LR cells, as well as their parental cells, detected by RT-PCR and western blot. E , F mRNA and protein levels of EZH2 in Huh7-LR and Hep3B-LR cells transfected with shEZH2#1, shEZH2#2, or shCtrl,, detected by RT-PCR and western blot. G , H Proliferation of both Huh7-LR and Hep3B-LR cells transfected with shEZH2#1 compared to those transfected with shCtrl at varying concentrations of lenvatinib. All data are presented as the means ± SDs, n = 5 per group, * ( p < 0.05), ** ( p < 0.01), ns = no significance

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: Expressing, CCK-8 Assay, Reverse Transcription Polymerase Chain Reaction, Western Blot, Transfection

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: EZH2 inhibits ferroptosis in HCC cells. A Analysis of cell death and apoptosis in Huh7-LR and Hep3B-LR cells after Lenvatinib (Lenv) treatment via Annexin V-FITC/PI staining. B Inhibitory ratio that was measured in Huh7-LR and Hep3B-LR using CCK-8 assay 24 h after treatment with Lenvatinib with or without Z-VAD-FMK, CQ, Nec-1 or Fer-1. C Proliferation of both Huh7-LR and Hep3B-LR cells transfected with shCtrl compared to those transfected with shEZH2#1 at varying concentrations of RSL3. D - F Relative levels of reactive oxygen species (ROS) (D), glutathione (GSH) ( E ), and malondialdehyde (MDA) ( F ) in Huh7-LR and Hep3B-LR cells transfected with shEZH2 #1 compared to shCtrl treated with Lenvatinib, RSL3 and Fer-1. (G-H) Intracellular lipid peroxidation and iron accumulation in Huh7-LR and Hep3B-LR cells transfected with shEZH2 #1 compared to shCtrl treated with Lenvatinib, RSL3 and Fer-1 were determined by C11-BODIPY and FerroOrange staining. All data are presented as the means ± SDs, n = 5 per group, * ( p < 0.05), ** ( p < 0.01), ns = no significance

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: Staining, CCK-8 Assay, Transfection

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: EZH2 suppression induced ferroptosis via enhancing ACSL1 expression through H3K27me3. A , B RT-qPCR assays demonstrated mRNA levels of indicated genes in in Huh7-LR (left panel) and Hep3B-LR (right panel) cells transfected with shEZH2 compared to shCtrl. C , D Expression levels of EZH2 in Huh7-LR ( C ) and Hep3B-LR ( D ) cells transfected with shEZH2 compared to shCtrl, detected by western blot. (E)Pearson correlation analysis of TCGA dataset revealed a significant negative correlation between EZH2 and ACSL1 expression. (F)Proliferation of Huh7-LR and Hep3B-LR cells transfected with shEZH2 or shACSL1. G - I Relative levels of ROS ( G ), GSH ( H ), and MDA ( I ) in Huh7-LR and Hep3B-LR cells transfected with shEZH2 or shACSL1. J-K Intracellular lipid peroxidation and iron accumulation in Huh7-LR and Hep3B-LR cells transfected with shEZH2 or shACSL1 were determined by C11-BODIPY and FerroOrange staining. L The protein levels of EZH2 and H3K27me3 in Huh7-LR cells transfected with shEZH2 or shCtrl were determined by western blot. M The enrichments of EZH2 and H3K27me3 on promoters of ACSL1 were determined by ChIP assays. N The altered enrichments of EZH2 and H3K27me3 on promoters of ACSL1 after EZH2 downregulatiton were determined by ChIP-qPCR assay. Normal IgG served as a negative control. All data are presented as the means ± SDs, n = 5 per group, * ( p < 0.05), ** ( p < 0.01), ns = no significance

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: Expressing, Quantitative RT-PCR, Transfection, Western Blot, Staining, ChIP-qPCR, Negative Control

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: EZH2 reduction enhances lenvatinib efficacy in treating resistant HCC in vivo. A Treatment schedule illustrating the timing of tumor inoculation and administration of treatments in BALB/c nude mice ( n = 6 per group). B , C Representative images of tumors ( B ) harvested on day 27 posttreatment, and corresponding average tumor weights ( C ) in each group. D Tumor growth curves representing the progression of Huh7-LR cell-induced tumors in mice treated with lenvatinib. E Expression levels of EZH2, H3K27me3 and ACSL1 in tumors. All data are presented as the means ± SDs, n = 6 per group, * ( p < 0.05), ** ( p < 0.01), ns = no significance

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: In Vivo, Expressing

Journal: BMC Cancer

Article Title: EZH2 confers lenvatinib resistance in hepatocellular carcinoma by suppressing ACSL1-Mediated ferroptosis

doi: 10.1186/s12885-025-15086-9

Figure Lengend Snippet: Inhibition of EZH2 enhances lenvatinib efficacy in treating hepatocellular carcinoma by promoting ACSL1-Mediated Ferroptosis via H3K27me3

Article Snippet: For immunostaining, tissue sections were incubated with a primary antibody targeting EZH2 (Cat# 21800-1-AP, Proteintech; working dilution 1:1000).

Techniques: Inhibition