Fig. 5: APR-246 repressed the expression of immune checkpoints and activated IFN signalling in p53 mutant cancer cells.
From: Mutant p53 gain of function mediates cancer immune escape that is counteracted by APR-246
a APR-246 repressed the expression of immune checkpoint molecules in cancer cell lines, as assessed by qPCR (n = 3). Human breast cancer cell lines: SKBR-3, MDAMB-231 and MDAMB-468. b APR-246 enhanced the expression of IFN signalling genes in cancer cell lines, as assessed by qPCR (n = 3). Human colorectal cancer cell line: DLD-1; human breast cancer cell lines: BT-549 and BT-20. c Induction by APR-246 of ERVs and IFN genes along with p53 target genes PMAIP1, TIGAR and SESN1 in HCT116 cells expressing p53 mutant R248W, as assessed by RT-qPCR (n = 3). d ERVs and IFN genes and p53 target genes PMAIP1, TIGAR and SESN1 are not significantly affected by APR-246 in p53-null HCT116 cells. e Induction by MDM2 inhibitor nutlin of ERVs and IFN genes along with p53 target genes PMAIP1, TIGAR and SESN1 in HCT116 cells expressing wtp53, as assessed by RT-qPCR (n = 3). f Comparison of basal levels of ERVs and IFN genes in isogenic HCT116 cells with different p53 status: wt, mutant and null, as assessed by RT-qPCR (n = 3). g Western blot assessment of the levels of p53 and MDM2 in wtp53 HCT116 cells treated/non-treated with nutlin (left panel), in HCT116p53R248W and HCT116p53null cells treated with APR-246 (middle panel) and in non-treated wtp53HCT116, p53null HCT116 and p53 mutant R248W-expressing HCT116 cells (right panel). a–e Each condition contains three biological repeats and each biological repeat includes three technical repeats. Unpaired Student’s t test, centre values are mean, error bars, s.d. p value is two-tailed. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. Treatment conditions were as follows: 12 h treatment of 40μM APR-246 on SKBR-3 and BT-549, 12 h treatment of 200μM APR-246 on MDAMB-231, 12 h treatment of 60 μM APR-246 on MDAMB-468, 24 h treatment of 90 μM APR-246 on DLD-1, 12 h treatment of 100 μM APR-246 on BT-20, 24 h treatment of 20 μM APR-246 on both HCT116p53R248W and HCT116p53null cells, 24 h treatment of 10 μM nutlin on wtp53HCT116 cells.
