Fig. 2
TP53 and RB1 inactivation drive CDC7 upregulation in lung and prostate adenocarcinoma. a CDC7 mRNA expression in adenocarcinoma clinical specimens, categorized by TP53/RB1 mutational status. Data obtained from LUAD TCGA (PanCancer, n = 237 wild type (wt), 33 mutated), LUAD OncoSG (n = 109 wt, 6 mutated)45 and PRAD SU2C/PCF Dream Team (n = 107 wt, 19 mutated)46 (b) Western blot images showing CDC7 protein expression in isogenic H1563 (LUAD) and 22PC (PRAD) cell lines with or without induced loss of function of TP53 and/or RB1 by shRNA against RB1 and dominant negative TP53 gene overexpression (H1563) or CRISPR/Cas9 knock out (22PC). Representative western blot image is shown (top) with quantification shown, as volume of the CDC7 band normalized by the volume of actin, and again normalized to control expression, which takes the value of 1. c Binding score for TP53 and E2F1 in the transcription start site of the CDC7 gene in different experimental settings including specimens from prostate and other tissues. Data obtained from The Signaling Pathways Project (ChIP-seq Atlas). d Barplot exhibiting data from CDC7 gene promoter reporter assays in isogenic H1563 (LUAD) and 22PC (PRAD) cell lines with or without induced inactivation of TP53 and/or and RB1, or with E2F1 overexpression. Normalized luciferase activity of a representative biological replicate is shown. e Barplot showing a representative biological replicate of an experiment assessing viability of control and TP53/RB1-inactivated H1563 (LUAD), 22PC and LnCap/AR (PRAD) cells treated with 0.5 µM simurosertib. Each condition shown was normalized to their matched untreated condition and represented as a normalized percentage for viability. For d and e, p-values were calculated using the Student’s t test (unpaired, heterogeneous variances, two-tailed). p-value legend: *<0.05, **<0.01, ***<0.001, ns, not significant
