Journal: Neoplasia (New York, N.Y.)

Article Title: Metabolic regulation of histone acetylation by ACLY supports MDR1 expression in colorectal cancer and highlights a targetable vulnerability

doi: 10.1016/j.neo.2026.101314

Figure Lengend Snippet: ACLY activity regulates MDR1 expression in colorectal cancer. (A) Transcript levels of ACLY and MDR1 (ABCB1) in colorectal cancer (red) and normal colon tissues (grey) analyzed using GEPIA (TCGA/GTEx datasets). (B) Immunoblot analysis of MDR1 in SW480 and DLD1 cells treated with the ACLY inhibitor BMS-303141 (20 or 50 μM, 48 h). (C) Immunoblot analysis of ACLY and MDR1 in SW480 and DLD1 cells transduced with empty vector (pLV) or ACLY-overexpressing vector (pLV[Exp]-hACLY). (D) Immunoblot analysis of ACLY and MDR1 in control and ACLY-overexpressing cells treated with BMS-303141 (50 μM, 48 h). (E) Relative ABCB1 mRNA levels in control and ACLY-overexpressing cells, and in cells treated with BMS-303141. (F) Relative ACLY mRNA levels under the same conditions. Data are presented as mean ± SD (n = 3 unless otherwise indicated). Statistical significance was determined using unpaired two-tailed t-tests or one-way ANOVA with Tukey’s post hoc test. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Human colorectal cancer cell lines SW480 (KRASG12V) and DLD1 (KRASG13D) were obtained from the American Type Culture Collection (ATCC) and authenticated by short tandem repeat profiling.

Techniques: Activity Assay, Expressing, Western Blot, Transduction, Plasmid Preparation, Control, Two Tailed Test

Journal: Neoplasia (New York, N.Y.)

Article Title: Metabolic regulation of histone acetylation by ACLY supports MDR1 expression in colorectal cancer and highlights a targetable vulnerability

doi: 10.1016/j.neo.2026.101314

Figure Lengend Snippet: ACLY activity modulates histone acetylation and MDR1 expression. (A) Immunoblot analysis of MDR1 in SW480 and DLD1 cells treated with the histone deacetylase inhibitor vorinostat (VOR; 0.5 μM for SW480 and 3.5 μM for DLD1) or DMSO for 24 h. Representative blots and densitometric quantification relative to control are shown. (B) Immunoblot analysis of acetylated histone H3 (H3K9ac) and histone H4 (H4K16ac) in SW480 wild-type (WT) and ACLY-overexpressing (OE) cells treated with vehicle or the ACLY inhibitor BMS-303141 (50 μM, 48 h). (C) Immunoblot analysis of H3K9ac and H4K16ac in DLD1 cells under the same conditions. β-actin was used as a loading control. Data are presented as mean ± SD (n = 3). Statistical significance was determined using unpaired two-tailed t-tests. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Human colorectal cancer cell lines SW480 (KRASG12V) and DLD1 (KRASG13D) were obtained from the American Type Culture Collection (ATCC) and authenticated by short tandem repeat profiling.

Techniques: Activity Assay, Expressing, Western Blot, Histone Deacetylase Assay, Control, Two Tailed Test

Journal: Neoplasia (New York, N.Y.)

Article Title: Metabolic regulation of histone acetylation by ACLY supports MDR1 expression in colorectal cancer and highlights a targetable vulnerability

doi: 10.1016/j.neo.2026.101314

Figure Lengend Snippet: ACLY expression is associated with resistance-related transcriptional programs in colorectal cancer. (A) Correlation analysis between ACLY expression and a gene set associated with lipid metabolism (ACLY, ACSS2, ACSS1, FASN, SREBP1) and drug transport pathways (ABCB1, ABCC2, ABCG5, EpCAM, CD24) in colorectal cancer samples using GEPIA2 (TCGA dataset). (B) Schematic representation of a proposed model linking ACLY-dependent acetyl-CoA production to histone acetylation and transcriptional regulation in CRC cells. (C) Relative mRNA expression of EpCAM, ABCC2, and CD24 in SW480 and DLD1 cells overexpressing ACLY compared with empty vector controls. (D) Relative mRNA expression of EpCAM, ABCC2, and CD24 in SW480 and DLD1 cells treated with the ACLY inhibitor BMS-303141 (50 μM) compared with vehicle-treated controls. Gene expression levels were determined by qPCR and normalized to ACTB. Data are presented as mean ± SEM (n = 3). Statistical significance was determined using unpaired two-tailed t-tests. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Human colorectal cancer cell lines SW480 (KRASG12V) and DLD1 (KRASG13D) were obtained from the American Type Culture Collection (ATCC) and authenticated by short tandem repeat profiling.

Techniques: Expressing, Drug Transport Assay, Plasmid Preparation, Gene Expression, Two Tailed Test

Journal: Neoplasia (New York, N.Y.)

Article Title: Metabolic regulation of histone acetylation by ACLY supports MDR1 expression in colorectal cancer and highlights a targetable vulnerability

doi: 10.1016/j.neo.2026.101314

Figure Lengend Snippet: Vitamin C induces coordinated changes in metabolic and chromatin-associated pathways in colorectal cancer cells. (A) Gene Ontology (GO) enrichment analysis of proteins differentially expressed following vitamin C treatment (5 mM, 4 h). (B) Volcano plot showing significantly upregulated and downregulated proteins (log₂ fold change > 1, p < 0.05). (C) KEGG pathway enrichment analysis highlighting pathways related to chromatin organization, DNA replication, nucleotide metabolism, and cell cycle regulation. (D) GO Cellular Component analysis showing enrichment of chromatin-associated complexes, including transcription regulator complexes, histone acetyltransferase-containing complexes, and Polycomb group (PcG) assemblies. (E) Heatmap representation of differentially expressed chromatin-associated proteins in control and vitamin C-treated cells. Proteomic analysis was performed in SW480 and DLD1 cells using label-free LC–MS/MS (diaPASEF). Data represent combined analysis of both cell lines.

Article Snippet: Human colorectal cancer cell lines SW480 (KRASG12V) and DLD1 (KRASG13D) were obtained from the American Type Culture Collection (ATCC) and authenticated by short tandem repeat profiling.

Techniques: Control, Liquid Chromatography with Mass Spectroscopy, Data-independent acquisition

Journal: Neoplasia (New York, N.Y.)

Article Title: Metabolic regulation of histone acetylation by ACLY supports MDR1 expression in colorectal cancer and highlights a targetable vulnerability

doi: 10.1016/j.neo.2026.101314

Figure Lengend Snippet: Metabolic and epigenetic consequences of vitamin C treatment in colorectal cancer cells. (A) Quantification of ¹³C-glucose-derived citrate in SW480 and DLD1 cells treated with vitamin C (5 mM) for 4 h (n = 3). (B) Immunoblot analysis of total ACLY and phosphorylated ACLY at Ser455 following vitamin C treatment (5 mM) (n = 3). (C) Immunoblot analysis and quantification of acetylated histone H4 (AcH4K16) and histone H3 (AcH3K9) in SW480 and DLD1 cells after vitamin C exposure (n = 3). (D) MDR1 (ABCB1) protein levels in SW480 and DLD1 cells treated with vitamin C (5 mM), quantified relative to vehicle control (n = 3). (E) Relative ACLY and ABCB1 mRNA expression determined by qPCR after 6 h of vitamin C treatment (5 mM) in SW480 and DLD1 cells (n = 3). Data are presented as mean ± SEM. Statistical significance was determined using unpaired two-tailed t-tests. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Human colorectal cancer cell lines SW480 (KRASG12V) and DLD1 (KRASG13D) were obtained from the American Type Culture Collection (ATCC) and authenticated by short tandem repeat profiling.

Techniques: Derivative Assay, Western Blot, Control, Expressing, Two Tailed Test

Journal: Journal of Cell Communication and Signaling

Article Title: Inflammatory cytokine IL‐6 regulates ADAMTS14 expression through MAPK and PI3K signaling in colorectal cancer

doi: 10.1002/ccs3.70092

Figure Lengend Snippet: Determination of the effect of IL‐6 application on ADAMTS14 expression. (A) Basal mRNA expression levels of the ADAMTS14 gene in different cancer cell lines were analyzed by qRT‐PCR. (B) SW480 cells were treated with 20 ng/mL IL‐6, and ADAMTS14 mRNA expression levels were determined by qRT‐PCR after the specified time periods (1, 6, 24, and 48 h). (C) ADAMTS14 protein levels after IL‐6 application were analyzed by Western blot, and β‐actin was used as a loading control. Protein band densities were analyzed densitometrically, and the fold change is shown in the graph. Western blot analysis was performed from a single experimental replicate; therefore, no error bars or SD values are presented. (D) ADAMTS14 protein expression was examined by immunofluorescence staining. Nuclei were stained with DAPI. Scale bar: 10 μm. All experiments were performed in triplicate, and data are presented as mean ± SD. Western blot analysis was performed from a single experimental replicate. Statistical analysis was performed using ANOVA, with * p < 0.05, ** p < 0.01, and *** p < 0.001 values. ANOVA, Analysis of Variance; DAPI, 4′,6‐Diamidino‐2‐Phenylindole; IL‐6, Interleukin‐6; qRT‐PCR, quantitative real‐time Polymerase Chain Reaction; SD, standard deviation.

Article Snippet: The human colorectal adenocarcinoma cell line SW480 (ATCC® CCL‐228TM, RRID:CVCL_0546) was used in this study.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Control, Immunofluorescence, Staining, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Journal of Cell Communication and Signaling

Article Title: Inflammatory cytokine IL‐6 regulates ADAMTS14 expression through MAPK and PI3K signaling in colorectal cancer

doi: 10.1002/ccs3.70092

Figure Lengend Snippet: Sequence analysis of the ADAMTS14 promoter region, promoter deletion constructions, and the effect of IL‐6 on promoter activity. (A) Multiple sequence alignment analysis of human, mouse, and rat ADAMTS14 promoter regions. Conserved nucleotide regions are shown in shaded form. (B) Base composition and CpG island analyses of the ADAMTS14 promoter region are shown. (C) Schematic representation of 5′ deletion constructions (−381/+297, −145/+297, and −43/+297) generated from the ADAMTS14 promoter region and cloning of these regions into the pMetLuc report vector. (D) Determination of the effect of IL‐6 (20 ng/mL) administration on ADAMTS14 promoter activity in SW480 cells by luciferase report analysis. Luciferase activity was normalized by SEAP. Data are presented as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001. (E) Schematic representation of predicted transcription factor binding motifs within the ADAMTS14 promoter region. The experimentally identified IL‐6‐responsive region (−145/−43 bp) is highlighted in gray. In silico promoter analysis identified multiple putative NF‐κB, AP‐1, SMAD2‐associated binding motifs clustered within this region, suggesting a transcriptionally active regulatory hotspot potentially involved in IL‐6‐responsive ADAMTS14 regulation. IL‐6, Interleukin‐6; NF‐κB, Nuclear Factor Kappa B; SEAP, Secreted Alkaline Phosphatase.

Article Snippet: The human colorectal adenocarcinoma cell line SW480 (ATCC® CCL‐228TM, RRID:CVCL_0546) was used in this study.

Techniques: Sequencing, Activity Assay, Generated, Cloning, Plasmid Preparation, Luciferase, Binding Assay, In Silico

Journal: Journal of Cell Communication and Signaling

Article Title: Inflammatory cytokine IL‐6 regulates ADAMTS14 expression through MAPK and PI3K signaling in colorectal cancer

doi: 10.1002/ccs3.70092

Figure Lengend Snippet: Identification of signaling pathways involved in IL‐6‐mediated ADAMTS14 regulation. (A) SW480 cells were treated with 20 ng/mL IL‐6 alone or in combination with different signaling pathway inhibitors (PD98059: ERK inhibitor, PD169316 : p38 MAPK, SP600125: JNK inhibitor, Wortmannin: PI3K inhibitor). ADAMTS14 mRNA expression levels were analyzed by Quantitative real‐time PCR after treatment. (B) Under the same conditions, ADAMTS14 protein levels were analyzed by Western blot, and β‐actin was used as a loading control. Protein band densities were analyzed densitometrical, and fold change is shown in the graph. (C) Luciferase report gene analysis was performed using the ADAMTS14 promoter construction containing the −43/+297 promoter region, and the effect of inhibitor administration in combination with IL‐6 on promoter activity was determined. Luciferase activity was calculated as the Luc/SEAP ratio. All experiments were performed in triplicate, and data are presented as mean ± standard deviation. Western blot analysis was performed from a single experimental replicate. Statistical analysis was performed using ANOVA, with * p < 0.05, ** p < 0.01, and *** p < 0.001. ANOVA, Analysis of Variance; ERK, Extracellular Signal‐Regulated Kinase; IL‐6, Interleukin‐6; MAPK, Mitogen‐Activated Protein Kinase; PI3K, Phosphatidylinositol 3‐Kinase; SEAP, Secreted Alkaline Phosphatase.

Article Snippet: The human colorectal adenocarcinoma cell line SW480 (ATCC® CCL‐228TM, RRID:CVCL_0546) was used in this study.

Techniques: Protein-Protein interactions, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Control, Luciferase, Activity Assay, Standard Deviation