Fig. 5: KDM5B promotes progression and cisplatin resistance in NPC cells through ZBTB16-mediated transcriptional inhibition of TOP2A.

a–e HNE1 and CNE2 cells were infected with indicated shZBTB16 or shTOP2A for 72 h. Cells were collected for Western blotting analysis (a), RT-qPCR analysis (b), colony-formation assay (c, only HNE1 cells) and transwell assay (d, only HNE1 cells) and MTS assay (e, HNE1 cells). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data presented as Mean ± SD with three replicates. NS not significant; *P < 0.05; **P < 0.01. ***P < 0.001. f HNE1 cells were infected with indicated shRNAs. After 72 h puromycin selection, cells were harvested and subcutaneously injected into nude mice for xenografts assay. Tumor volumes were measured every 3 days. Tumors were harvested, photographed, and weighed. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data presented as Mean ± SD with five replicates. NS not significant; *P < 0.05; **P < 0.01. g, h HNE1 and CNE2 cells were infected with indicated shKDM5B or shZBTB16 for 72 h. Cells were collected for Western blotting analysis (g), RT-qPCR analysis (h). Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data presented as Mean ± SD with three replicates. NS not significant; *P < 0.05; **P < 0.01. ***P < 0.001. i HNE1 and CNE2 cells were infected with shControl or shTOP2A plasmid for 48 h. then cells were treated with or without cisplatin (4 μg/ml) for another 48 h. Cells were collected for CCK-8 assay. j HNE1 and CNE2 cells were infected with indicated shRNAs for 72 h. Cells were treated with a serial dose of cisplatin. Then, these cells were collected for CCK-8 assay and subjected to measure the IC₅₀ values of cisplatin. k HNE1 and CNE2 cells were infected with indicated shRNAs for 48 h. then cells were treated with or without cisplatin (4 μg/ml) for another 48 h. Cells were collected for colony-formation assay. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test. Data presented as Mean ± SD with three replicates. NS not significant; *P < 0.05; **P < 0.01; ***P < 0.001.