Fig. 5: Increased MEK/ERK pathway activity and dependency in aneuploid cells.
From: Human aneuploid cells depend on the RAF/MEK/ERK pathway for overcoming increased DNA damage
a Western Blot of pMEK1/2 (Ser217/221) and MEK1/2 in pseudo-diploid and highly-aneuploid clones. b Quantification of MEK1/2 activation (pMEK/MEK ratio). n = 5 independent experiments; *p = 0.0383 (SS51), p = 0.0247 (SS111); two-sided One Sample t-test. c Western Blot of pERK1/2 (Thr202/Tyr204) and ERK1/2 in pseudo-diploid and highly-aneuploid clones. d Quantification of ERK1/2 activation (pERK/ERK ratio). n = 5 independent experiments; *p = 0.0346 (SS51) and p = 0.0223 (SS111); two-sided One Sample t-test. e Sensitivity to 72 h drug treatment with the MEK inhibitor trametinib, in pseudo-diploid clones vs. highly-aneuploid clones. Fold-change calculated relative to SS48. n = 5 independent experiments; ****p = 6.6*10−5 and p = 1.9*10−5 for SS51 and SS111 respectively; two-sided one-sample t-test. f Sensitivity to 72 h drug treatment with the ERK inhibitor ulixertinib, in pseudo-diploid vs. highly-aneuploid clones. Fold-change calculated relative to SS48. n = 5 independent experiments; **p = 0.0011 (SS31) and p = 0.0016 (SS111), ***p = 0.0007 (SS51); two-sided one-sample t-test. g Western blot of pMEK1/2 and total MEK1/2 in reversine-pulsed RPE1 cells. h Quantification of MEK activation in the reversine-pulsed cells, relative to the DMSO control. n = 6 independent experiments. **p = 0.0096; two-sided one-sample t-test. i Western blot of pERK1/2 and total ERK1/2 in reversine-pulsed RPE1 cells. j Quantification of ERK activation in reversine-treated cells, relative to the DMSO control. n = 6 independent experiments. *p = 0.0148; two-sided one-sample t-test. k, l Comparison of MEK (k) and ERK (l) activity between the top and bottom aneuploidy quartiles of human cancer cell lines (n = 460). Data obtained from DepMap 22Q1 release.***p = 0.0006 (MEK); *p = 0.0424 (ERK); two-tailed Mann–Whitney test. m Sensitivity to the MEK inhibitor trametinib in the top and bottom aneuploidy quartiles of human cancer cell lines (n = 412). Data obtained from GDSC1 drug screen 22Q1 release. **p = 0.0069; two-tailed Mann-Whitney test. n PRISM-based comparison of drug sensitivity to 120 h treatment with the MEK inhibitor selumetinib, between cancer cells treated with reversine (250 nM) or with control DMSO (n = 84). ****p = 5.3*10−9; two-tailed Wilcoxon rank sum test. o Drug sensitivity to 72 h etoposide treatment, in RPE1 and MEK-overexpressing (MEK OE) RPE1 cells. Fold-change calculated relative to RPE1, per experiment. n = 5 independent experiments; *p = 0.0256; two-sided one-sample t-test. p Viability following 72 h treatment with a sub-lethal dose (0.45 nM) of the MEK inhibitor trametinib or DMSO, in combination with etoposide (2.5 µM) in highly-aneuploid clones. n = 5 independent experiments. Fold change calculated relative to etoposide-treated cells, per experiment; ***p = 0.0009 (SS51) and **p = 0.0015 (SS111); two-sided one-sample t-test. q Comparison of the RAF/MEK/ERK pathway activity by ssGSEA in pancreatic PDXs (GSE235843) treated with olaparib. *p = 0.036; one-tailed Mann–Whitney test. r Comparison of the RAF/MEK/ERK pathway activity by ssGSEA in breast tumors (GSE173839) treated with olaparib in combination with durvalumab. **p = 0. 006; one-tailed Mann–Whitney test. Source data are provided as a Source Data file.
