10.6084/m9.figshare.4754500
Peter John Webster
Peter John
Webster
Anna Tiffany Littlejohns
Anna Tiffany
Littlejohns
Hannah Jane Gaunt
Hannah Jane
Gaunt
K. Raj Prasad
K. Raj
Prasad
David John Beech
David John
Beech
Dermot Anthony Burke
Dermot Anthony
Burke
AZD1775 induces toxicity through double-stranded DNA breaks independently of chemotherapeutic agents in p53-mutated colorectal cancer cells
Taylor & Francis Group
2017
colorectal cancer
checkpoint kinase
cell cycle
DNA damage
mitosis
2017-11-09 09:31:27
Dataset
https://tandf.figshare.com/articles/dataset/AZD1775_Induces_Toxicity_Through_Double-Stranded_DNA_Breaks_Independently_of_Chemotherapeutic_Agents_in_p53-Mutated_Colorectal_Cancer_Cells/4754500
<p>AZD1775 is a small molecule WEE1 inhibitor used in combination with DNA-damaging agents to cause premature mitosis and cell death in p53-mutated cancer cells. Here we sought to determine the mechanism of action of AZD1775 in combination with chemotherapeutic agents in light of recent findings that AZD1775 can cause double-stranded DNA (DS-DNA) breaks. AZD1775 significantly improved the cytotoxicity of 5-FU in a p53-mutated colorectal cancer cell line (HT29 cells), decreasing the IC<sub>50</sub> from 9.3 μM to 3.5 μM. Flow cytometry showed a significant increase in the mitotic marker pHH3 (3.4% vs. 56.2%) and DS-DNA break marker γH2AX (5.1% vs. 50.7%) for combination therapy compared with 5-FU alone. Combination therapy also increased the amount of caspase-3 dependent apoptosis compared with 5-FU alone (4% vs. 13%). The addition of exogenous nucleosides to combination therapy significantly rescued the increased DS-DNA breaks and caspase-3 dependent apoptosis almost to the levels of 5-FU monotherapy. In conclusion, AZD1775 enhances 5-FU cytotoxicity through increased DS-DNA breaks, not premature mitosis, in p53-mutated colorectal cancer cells. This finding is important for designers of future clinical trials when considering the optimal timing and duration of AZD1775 treatment.</p>