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(A) <t>Methylation</t> index, calculated as the ratio of SAM to SAH), of OVCAR3 across a time course of platinum (cisplatin, CDDP) treatment (3, 8 and 16 hours) (B) Global 5-methylcytosine (5-mC) levels of OVCAR3 cells untreated (UT) or treated with CDDP (15μM, 16 hours) measured by ELISA and normalized to total level of <t>DNA.</t> (C) ssGSEA-derived ΔDNMT1 (left) and ΔDNMT3A (right) scores plotted against ΔMAT2A expression between matched recurrent and primary tumors stratified by MAT2A-high (red) and MAT2A-low (blue) patients. (D) Heatmap of beta values for statistically significant (adjusted p<0.05) differentially methylated probes identified between siScramble and CDDP-treated conditions. (E) Volcano plot of CpG-level methylation changes between CDDP-treated and siScramble control cells. Each point represents a single CpG site; blue points indicate statistically significant differential β-value (adjusted p < 0.05, Δβ> 0.05); red points indicate β-value changes below statistical significance; yellow points indicate statistically significant but Δβ-value < 0.05. Positive and negative β-value denote hypermethylation and hypomethylation in CDDP versus siScramble, respectively. (F) Pie chart of the distribution of hypermethylated CpG to gene features (promoter, exon, intron, and intergenic regions) in CDDP versus siScramble. (G) Pie chart of genomic annotation of differentially methylated regions (DMRs) across CpG island contextual categories (island, shore, shelf, and open sea) in CDDP versus siScramble. (H) Ingenuity Pathway Analysis (IPA) of biological pathways enriched among DMR-associated genes identified from the CDDP versus siScramble. (* p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001; ns, not significant)
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(A) <t>Methylation</t> index, calculated as the ratio of SAM to SAH), of OVCAR3 across a time course of platinum (cisplatin, CDDP) treatment (3, 8 and 16 hours) (B) Global 5-methylcytosine (5-mC) levels of OVCAR3 cells untreated (UT) or treated with CDDP (15μM, 16 hours) measured by ELISA and normalized to total level of <t>DNA.</t> (C) ssGSEA-derived ΔDNMT1 (left) and ΔDNMT3A (right) scores plotted against ΔMAT2A expression between matched recurrent and primary tumors stratified by MAT2A-high (red) and MAT2A-low (blue) patients. (D) Heatmap of beta values for statistically significant (adjusted p<0.05) differentially methylated probes identified between siScramble and CDDP-treated conditions. (E) Volcano plot of CpG-level methylation changes between CDDP-treated and siScramble control cells. Each point represents a single CpG site; blue points indicate statistically significant differential β-value (adjusted p < 0.05, Δβ> 0.05); red points indicate β-value changes below statistical significance; yellow points indicate statistically significant but Δβ-value < 0.05. Positive and negative β-value denote hypermethylation and hypomethylation in CDDP versus siScramble, respectively. (F) Pie chart of the distribution of hypermethylated CpG to gene features (promoter, exon, intron, and intergenic regions) in CDDP versus siScramble. (G) Pie chart of genomic annotation of differentially methylated regions (DMRs) across CpG island contextual categories (island, shore, shelf, and open sea) in CDDP versus siScramble. (H) Ingenuity Pathway Analysis (IPA) of biological pathways enriched among DMR-associated genes identified from the CDDP versus siScramble. (* p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001; ns, not significant)
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(A) <t>Methylation</t> index, calculated as the ratio of SAM to SAH), of OVCAR3 across a time course of platinum (cisplatin, CDDP) treatment (3, 8 and 16 hours) (B) Global 5-methylcytosine (5-mC) levels of OVCAR3 cells untreated (UT) or treated with CDDP (15μM, 16 hours) measured by ELISA and normalized to total level of <t>DNA.</t> (C) ssGSEA-derived ΔDNMT1 (left) and ΔDNMT3A (right) scores plotted against ΔMAT2A expression between matched recurrent and primary tumors stratified by MAT2A-high (red) and MAT2A-low (blue) patients. (D) Heatmap of beta values for statistically significant (adjusted p<0.05) differentially methylated probes identified between siScramble and CDDP-treated conditions. (E) Volcano plot of CpG-level methylation changes between CDDP-treated and siScramble control cells. Each point represents a single CpG site; blue points indicate statistically significant differential β-value (adjusted p < 0.05, Δβ> 0.05); red points indicate β-value changes below statistical significance; yellow points indicate statistically significant but Δβ-value < 0.05. Positive and negative β-value denote hypermethylation and hypomethylation in CDDP versus siScramble, respectively. (F) Pie chart of the distribution of hypermethylated CpG to gene features (promoter, exon, intron, and intergenic regions) in CDDP versus siScramble. (G) Pie chart of genomic annotation of differentially methylated regions (DMRs) across CpG island contextual categories (island, shore, shelf, and open sea) in CDDP versus siScramble. (H) Ingenuity Pathway Analysis (IPA) of biological pathways enriched among DMR-associated genes identified from the CDDP versus siScramble. (* p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001; ns, not significant)
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(A) Methylation index, calculated as the ratio of SAM to SAH), of OVCAR3 across a time course of platinum (cisplatin, CDDP) treatment (3, 8 and 16 hours) (B) Global 5-methylcytosine (5-mC) levels of OVCAR3 cells untreated (UT) or treated with CDDP (15μM, 16 hours) measured by ELISA and normalized to total level of DNA. (C) ssGSEA-derived ΔDNMT1 (left) and ΔDNMT3A (right) scores plotted against ΔMAT2A expression between matched recurrent and primary tumors stratified by MAT2A-high (red) and MAT2A-low (blue) patients. (D) Heatmap of beta values for statistically significant (adjusted p<0.05) differentially methylated probes identified between siScramble and CDDP-treated conditions. (E) Volcano plot of CpG-level methylation changes between CDDP-treated and siScramble control cells. Each point represents a single CpG site; blue points indicate statistically significant differential β-value (adjusted p < 0.05, Δβ> 0.05); red points indicate β-value changes below statistical significance; yellow points indicate statistically significant but Δβ-value < 0.05. Positive and negative β-value denote hypermethylation and hypomethylation in CDDP versus siScramble, respectively. (F) Pie chart of the distribution of hypermethylated CpG to gene features (promoter, exon, intron, and intergenic regions) in CDDP versus siScramble. (G) Pie chart of genomic annotation of differentially methylated regions (DMRs) across CpG island contextual categories (island, shore, shelf, and open sea) in CDDP versus siScramble. (H) Ingenuity Pathway Analysis (IPA) of biological pathways enriched among DMR-associated genes identified from the CDDP versus siScramble. (* p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001; ns, not significant)

Journal: bioRxiv

Article Title: Targeting MAT2A-S-adenosylmethionine (SAM) Axis Attenuates DNA Damage Response and Cancer Stemness to Increase Platinum Sensitivity in Ovarian Cancer

doi: 10.64898/2026.09.22.753550

Figure Lengend Snippet: (A) Methylation index, calculated as the ratio of SAM to SAH), of OVCAR3 across a time course of platinum (cisplatin, CDDP) treatment (3, 8 and 16 hours) (B) Global 5-methylcytosine (5-mC) levels of OVCAR3 cells untreated (UT) or treated with CDDP (15μM, 16 hours) measured by ELISA and normalized to total level of DNA. (C) ssGSEA-derived ΔDNMT1 (left) and ΔDNMT3A (right) scores plotted against ΔMAT2A expression between matched recurrent and primary tumors stratified by MAT2A-high (red) and MAT2A-low (blue) patients. (D) Heatmap of beta values for statistically significant (adjusted p<0.05) differentially methylated probes identified between siScramble and CDDP-treated conditions. (E) Volcano plot of CpG-level methylation changes between CDDP-treated and siScramble control cells. Each point represents a single CpG site; blue points indicate statistically significant differential β-value (adjusted p < 0.05, Δβ> 0.05); red points indicate β-value changes below statistical significance; yellow points indicate statistically significant but Δβ-value < 0.05. Positive and negative β-value denote hypermethylation and hypomethylation in CDDP versus siScramble, respectively. (F) Pie chart of the distribution of hypermethylated CpG to gene features (promoter, exon, intron, and intergenic regions) in CDDP versus siScramble. (G) Pie chart of genomic annotation of differentially methylated regions (DMRs) across CpG island contextual categories (island, shore, shelf, and open sea) in CDDP versus siScramble. (H) Ingenuity Pathway Analysis (IPA) of biological pathways enriched among DMR-associated genes identified from the CDDP versus siScramble. (* p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001; ns, not significant)

Article Snippet: For methylation analysis, 500 ng of DNA was bisulfite-converted using the EZ DNA Methylation Kit (Zymo Research, cat. D5031) following the manufacturer’s instructions.

Techniques: Methylation, Platinum, Enzyme-linked Immunosorbent Assay, Derivative Assay, Expressing, Control, Analysis, Significance Assay