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Determination of DNA lesion bypass using a ChIP-based assay
Dayong Wu1, Ananya Banerjee1, Shurui Cai1
1Department of Radiation Oncology, The Ohio State University, Columbus, OH, 43210, USA; Comprehensive Cancer Center, The Ohio State University, Columbus, OH, 43210, USA.
DNA Repair
|September 27, 2021
Summary
We developed a new method to measure DNA lesion bypass in cancer cells. This technique reveals that ovarian cancer stem cells have higher DNA lesion bypass activity, impacting chemoresistance.
Area of Science:
- Molecular Biology
- Genetics
- Cancer Research
Background:
- DNA lesion bypass is crucial for DNA damage tolerance and cell survival.
- Understanding lesion bypass mechanisms is key to addressing carcinogenesis and chemoresistance.
Purpose of the Study:
- To develop and validate a novel chromatin immunoprecipitation (ChIP)-based assay for measuring DNA lesion bypass activity.
- To assess lesion bypass efficiency across cisplatin-induced DNA crosslinks in cancer cells.
Main Methods:
- Developed a ChIP-based assay to detect and quantify newly synthesized DNA (BrdU-labeled) opposite cisplatin-induced intrastrand crosslinks.
- Validated the assay using ovarian cancer cells with inhibited translesion synthesis (TLS) activity.
Main Results:
- The ChIP assay successfully measured lesion bypass activity across cisplatin-induced DNA lesions.
- Ovarian cancer stem cells demonstrated significantly higher lesion bypass activity compared to bulk cancer cells from the same line.
Conclusions:
- The novel ChIP-based assay effectively quantifies DNA lesion bypass in live cells.
- This method provides insights into cancer stem cell behavior and may be adaptable for studying other DNA lesions.
Keywords:
Cancer stem cellChIPChromatin immunoprecipitationCisplatinLesion bypassTLSTranslesion synthesis
