Journal: iScience
Article Title: Identification of ADAR1i-124: The first effective A-to-I RNA editing inhibitor with promising cancer therapeutic potential
doi: 10.1016/j.isci.2025.114615
Figure Lengend Snippet: DNA methylation inhibitor 5-Aza-CdR enhances the efficacy of ADAR1i-124 in dose-dependent eradication of certain cancer cell lines less responsive to ADAR1i-124 alone (A) 5-Aza-CdR (1 μM) reduced the IC 50 of ADAR1i-124 for Yumm1.7 CM cell survival rate by 12-fold. A similar reduction in cell viability was observed with the combination treatment of 5-Aza-CdR in A172 cells, which are completely unresponsive to ADAR1i-124 monotherapy. 5-Aza-CdR (1 μM) alone had no significant effect on the viability of either Yumm1.7 or A172 cells (left graphs). Data: mean ± SD ( n = 3, biological replicates). Significant differences were identified by two-tailed Student’s t tests: n.s., not significant. (B) Immunostaining with J2 antibodies revealed unedited and/or under-edited dsRNAs in Yumm1.7 cells treated with just 1 μM of ADAR1i-124, compared to the previously used 10 μM concentration (see B). These unedited and/or under-edited dsRNAs were detected also in A172 cells treated with the combination of ADAR1i-124 (10 μM) and 5-Aza-CdR (1 μM). Scale bars, 20 μm. (C) Immunostaining of Z-RNAs with Z22 antibodies in Yumm1.7 cells treated with 5-Aza-CdR and/or ADAR1i-124 (1 μM). Scale bars, 20 μm. (D) Western blotting analysis for ADAR1 downstream targets in Yumm1.7 and A172 cells treated with ADAR1i-124 and/or 5-Aza-CdR. Apparent molecular weights (kDa) are indicated. Full blots with molecular weight markers are shown in H (Yumm1.7) and I (A172). (E) The decrease in the Yumm1.7 cell survival rate by a combination of 5-Aza-CdR and ADAR1i-124 was rescued by siIfih1 (MDA5) and siZbp1, while the inhibition of the A172 cell survival rate by a combination of 5-Aza-CdR and ADAR1i-124 was rescued by siIFIH1 (MDA5) and siEIF2ak2 (PKR). Data: mean ± SD ( n = 3 per group, biological replicates). One-way ANOVA followed by Tukey’s post hoc test was used to determine the significance. n.s., not significant; ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001.
Article Snippet: HeLa human ovarian carcinoma (ATCC CCL-2), IMR90 human lung fibroblast (ATCC CCL-186), WM3000 and WM4223 human acral melanoma, HeLa-Nluc-edit cells, A172 human glioblastoma, Yumm1.7 mouse cutaneous melanoma (ATCC CRL-3362), and HGS2 mouse ovarian cancer cell lines were used in this study.
Techniques: DNA Methylation Assay, Two Tailed Test, Immunostaining, Concentration Assay, Western Blot, Molecular Weight, Inhibition
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