Fig. 4

RNAi-mediated silencing of endogenous Cirbp remarkably enhanced the tumor-killing effect of hyperthermia on NPC cells and cancer stem-like cells in vitro. A qRT-PCR assay for detecting the expression of Cirbp, Rbm3, miR-143 and miR-142-5p in the indicated NPC cells treated with or without hyperthermia at 42 °C for 30 min. B Western blot was employed to detect Cirbp expression in CNE2, SUNE1 and HONE1-EBV cells treated with or without hyperthermia at 42 °C for 30 min. C Class comparison and hierarchical clustering of differentially expressed hyperthermia-related genes between NPC cells treated with or without hyperthermia at 42 °C for 30 min. A cluster heat map for upregulated (red) and downregulated (blue) genes (see Tables S3 and S4) is shown. Other details as in Fig. S5. D Gene ontology (GO) analysis of up- and down-regulated genes (see Table S3) enriched in hyperthermia-associated biological processes, such as cellular response to heat, DNA damage and repair, cell cycle and cell death between NPC cells treated with or without hyperthermia at 42 °C for 30 min. E–H CCK-8 assay E, colony formation assay F, tumor sphere formation assay G and AnnexinV/PI apoptosis assay H were performed in shSCR- or shCirbp-expressing NPC cells treated with or without hyperthermia at 42 °C for 30 min. SCR: scrambled control shRNA. I Heatmap showing selected differentially expressed genes (see Table S6) related to cell death in shCirbp-expressing NPC cells. Right column lists the selected gene symbols. J GO and KEGG pathway analysis of up- and down-regulated genes (see Tables S6 and S7) related to cell survival and death in shSCR and shCirbp-expressing NPC cells