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. 2024 Jan 26;5(2):101393. doi: 10.1016/j.xcrm.2024.101393

Figure 5.

Figure 5

The direct effects of cisplatin versus carboplatin on downstream immune-related programs in cancer cells are mediated by the DNA damage transducer ATR

(A) Enrichment of Hallmark gene sets with cisplatin (red) versus carboplatin (gray) treatment in the 5637 human bladder cancer cell line (top), RT112 human bladder cancer cell line (middle), and MC38 murine colon cancer cell line (bottom). The cisplatin and carboplatin concentrations used were 5 and 35 μM, respectively, and cells were collected 24 h after treatment. Only pathways that were significantly changed with cisplatin versus carboplatin (false discovery rate [FDR] < 0.05) are shown. n = 3 per treatment group for the 5637 and RT112 cell lines; n = 2 per treatment group for the MC38 cell line.

(B) Protein expression of PD-L1 as determined by the median fluorescence intensity in the three cell lines treated with increasing concentrations of cisplatin (red) or carboplatin (black) for 24 h. Data depict the aggregate of three independent experiments (mean ± SEM).

(C) Representative immunoblots showing p-ATM S1981, p-ATR T1989, p-Chk1 S317, p-Chk2 T68, and GAPDH levels in lysates of the 5637 cell line treated with increasing concentrations of cisplatin or carboplatin for 6 h. One representative experiment out of four independent experiments is shown.

(D) Protein expression of PD-L1 as determined by median fluorescence intensity in the 5637 cell line treated with increasing concentrations of cisplatin or carboplatin for 24 h, in the presence or absence of an ATM inhibitor (KU-55933, 1 μM) or ATR inhibitor (VE-821, 1 μM). Data depict the aggregate of three independent experiments (mean ± SEM).

(E and F) Bulk RNA-seq analysis of 5637 cells treated with 5 μM cisplatin or 35 μM carboplatin, in the absence (gold) or presence of 1 μM ATR inhibitor (navy blue) for 24 h.

(E) Enrichment of Hallmark gene sets. n = 4 per treatment group.

(F) Heatmap displaying the scaled transcriptional expression of genes involved in immune-related transcriptional programs. Heatmap shows an average score of n = 4 individual samples per treatment group. See also Figures S5–S8.