Research Paper Volume 13, Issue 12 pp 15833—15874

The lack of functional DNMT2/TRDMT1 gene modulates cancer cell responses during drug-induced senescence

DNMT2/TRDMT1 gene knockout-mediated apoptosis and necrosis in U-251 MG glioblastoma cells treated with DOX or ETOPO for 24 h and AZA post-treatment for 24 h and immediately assayed (A) and assayed after 7 days of drug removal and AZA post-treatment for 24 h (B). Apoptosis and necrosis were analyzed using flow cytometry. Representative dot plots are shown. Bars indicate SD, n = 3, ***p **p *p a posteriori test), ###p a posteriori test). CTR, control conditions; DOX, doxorubicin treatment; ETOPO, etoposide treatment; AZA, azacytidine treatment; MIX, azacytidine post-treatment; C-NIC, control cells with unmodified levels of DNMT2/TRDMT1 containing control plasmid; D-NIC, cells with DNMT2/TRDMT1 gene knockout containing dedicated DNMT2 double nickase plasmid.

Figure 3. DNMT2/TRDMT1 gene knockout-mediated apoptosis and necrosis in U-251 MG glioblastoma cells treated with DOX or ETOPO for 24 h and AZA post-treatment for 24 h and immediately assayed (A) and assayed after 7 days of drug removal and AZA post-treatment for 24 h (B). Apoptosis and necrosis were analyzed using flow cytometry. Representative dot plots are shown. Bars indicate SD, n = 3, ***p < 0.001, **p < 0.01, *p < 0.05 compared to CTR (ANOVA and Dunnett’s a posteriori test), ###p < 0.001 compared to drug-treated C-NIC cells (ANOVA and Tukey’s a posteriori test). CTR, control conditions; DOX, doxorubicin treatment; ETOPO, etoposide treatment; AZA, azacytidine treatment; MIX, azacytidine post-treatment; C-NIC, control cells with unmodified levels of DNMT2/TRDMT1 containing control plasmid; D-NIC, cells with DNMT2/TRDMT1 gene knockout containing dedicated DNMT2 double nickase plasmid.