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Fig. 3 | Molecular Cancer

Fig. 3

From: Aberrant m5C hypermethylation mediates intrinsic resistance to gefitinib through NSUN2/YBX1/QSOX1 axis in EGFR-mutant non-small-cell lung cancer

Fig. 3

NSUN2 deficiency overcomes intrinsic gefitinib resistance in vitro and in vivo. a H1650 and H1975 cells transfected with siRNA targeting NSUN2 (siNSUN2) or non-targeting control (siCtrl) were exposed to gefitinib (1 µM) for 72 h and cell viability was measured by CCK-8 assay. NSUN2 knockdown efficacy was evaluated using western blotting (top panel). b H1650 and H1975 cells transfected with siNSUN2 or siCtrl were exposed to gefitinib (1 µM) for 72 h and cell apoptosis was detected by Annexin V-FITC/PI staining. c H1650 cells transfected with siNSUN2 or shNSUN2 were detected for EGFR protein expression by western blotting analysis. d, e H1650 and H1975 cells pre-treated with shNSUN2 were stably transfected with NSUN2-WT or NSUN2-DM and cell proliferation was detected by CCK-8 (d) or colony formation assay (e). Rescue efficacy of NSUN2 was evaluated by western blotting (right panel). f, g Mean volumes (f) and tumor weights (g) of tumor xenografts obtained from BALB/c nude mice subcutaneously implanted with H1650-shCtrl or H1650-shNSUN2 cells (n = 7 per group). h, i Growth curve (h) and average tumor weights (i) of H1975-shCtrl and H1975-shNSUN2-derived xenografts in the subcutaneous implantation mouse model (n = 6 per group). Error bars represent means ± SD. For (h), results were expressed as mean ± SEM. For a-e, n = 3 biological independent experiments. ns, not significant, *p < 0.05, **p < 0.01, ***p < 0.001.

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