FIGURE 1 from DNA2 Nuclease Inhibition Confers Synthetic Lethality in Cancers with Mutant p53 and Synergizes with PARP Inhibitors
Year: 2023
Type: other
Abstract: DNA2 is upregulated in many types of cancer, which is at least in part mediated by mutant p53. A, Expression of DNA2 is significantly elevated in 14 cancers from TCGA database compared with normal tissues. Tumor samples are denoted with bolded lines. Significance is calculated by Mann–Whitney U test. AC, adenocarcinoma; SC, squamous cell carcinoma; CC, clear cell; CS, carcinosarcoma; EC, endometrial carcinoma. B, Expression of DNA2 in breast invasive carcinoma, lung squamous carcinoma, uterine corpus endometrial carcinoma, and ovarian serous carcinoma from TCGA dataset based on TP53 mutation status. The data from other types of cancer are shown in Supplementary Fig. S1. WT: wild type. C, Depletion of mutp53 decreased DNA2 mRNA expression in MDA-MB-231 and MDA-MB-468 cells. Gene expression data were extracted from GSE26262 (32) and GSE31812 (33), in which gene expression was performed using Affymetrix microarray and normalized using the Robust Multichip Average (RMA) algorithm (34). In MDA-MB-468 cells, the short hairpin RNA (shRNA) against mutp53 was induced by doxycycline (Dox). D, DNA2 mRNA and protein expression in p53-null H1299 cells transfected with either an empty vector, WT p53, or indicated mutp53 (V143A, R175H, R248W, R249S, and R273H). Forty-eight hours after transfection, DNA2 mRNA was measured by qRT-PCR and normalized by GAPDH. *, P < 0.05; **, P < 0.01; and ***, P < 0.001 indicating statistically significant difference (two-tailed t test). Right: p53 and DNA2 immunoblots. The relative intensities of DNA2 protein were quantified by NIH ImageJ software, and normalized by loading control GAPDH. E, Depletion of mutant p53 decreases DNA2 protein levels. MDA-MB-468, MDAH-2774 or BT549 cells stably expressing either a scrambled shRNA or a mutp53 shRNA were harvested and the total cell lysates were subjected to immunoblotting using antibodies specific to DNA2, p53 or GAPDH. The relative intensities of DNA2 were quantified by NIH ImageJ software, and normalized by loading control GAPDH. The right panel shows the fold difference of DNA2 protein in MDA-MB-468 (n = 3 biological replicates), MDAH-2774 (n = 4 biological replicates), or BT549 (n = 2 biological replicates) stably expressing a scrambled shRNA versus a mutp53 shRNA. F, Box plots of DNA2 mRNA expression in chemotherapy responder or nonresponder patients (defined by relapse-free survival at 6 months) of serous ovarian cancer. Data were analyzed from TCGA dataset using ROC plotter platform. Left: all chemotherapy; middle: platin; right: taxane.
Language: English
Subfield: Oncology
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W4387673814