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Updated: Mar 1, 2026
Visualizing DNA Damage Repair Proteins in Patient-Derived Ovarian Cancer Organoids via Immunofluorescence Assays
Published on: February 24, 2023
[Functional Analysis of DNA Damage Repair Factor WDR70 and Its Mutation in Ovarian Cancer]
Lian-di Guo1, Dan Wang1, Fan Yang1
1Southwest University for Nationalities, Chengdu 610041, China.
Objectives:
To analyze the cellular function of the newly discovered DNA damage repair factor WDR70, and investigate the mutation in ovarian cancer to verify if function loss of the WDR70gene was associated with ovarian cancer.
Methods:
The WDR70 gene was silenced by using siRNA technique or overexpressed its wild and mutation type by with lentivirus and plasmid in hunman cells. The subcellular localization and biochemical function of WDR70 was analyzes by indirect immunofluorescence and immunoblotting. The expression level of WDR70 and the mutations of its cDNA was checked with RT-PCR sequencing for 1 normal ovarian tissue and 16 ovarian cancer specimen.
Results:
We found gene silencing of WDR70 or overexpression of WDR70 mutation type disrupts the phosphorylation level of homologous recombination functional protein RPA32 and the ability of recruitment at DNA damage site of recombinase RAD51, the loss of function of WDR70 also causes the elevation of the chromosome breakage in metaphase. Meanwhile, we also noticed that the existence of multiple mutations in genomic WDR70 in ovarian cancer specimen.
Conclusions:
Our results defined that in vitro system, WDR70 is a DNA damage repair gene, silencing of WDR70 or overexpression of WDR70 mutation type disrupts homologous recombination and chromosomal instability; the frequent mutations of WDR70 gene in genome of ovarian cancer specimens could also lead to DNA repair defeat and gene instability. Consequently WDR70 gene could represent an anti-cancer mechanism for ovarian cancer.
Insights
The DNA damage repair gene WDR70 is crucial for homologous recombination and chromosomal stability. Mutations in WDR70 are linked to ovarian cancer, suggesting its role in DNA repair and as an anti-cancer mechanism.
Area of Science:
- Molecular Biology
- Genetics
- Oncology
Background:
- WDR70 is a newly identified factor involved in DNA damage repair.
- Understanding its cellular function and role in cancer is essential.
Purpose of the Study:
- To elucidate the cellular function of WDR70 in DNA repair.
- To investigate WDR70 mutations in ovarian cancer and their association with disease.
Main Methods:
- Gene silencing (siRNA) and overexpression (lentivirus, plasmid) of WDR70 in human cells.
- Analysis of subcellular localization and biochemical function using immunofluorescence and immunoblotting.
- RT-PCR sequencing of WDR70 in ovarian cancer tissues.
Main Results:
- WDR70 dysfunction disrupts homologous recombination protein phosphorylation (RPA32) and RAD51 recruitment.
- Loss of WDR70 function leads to increased chromosomal breakage.
- Multiple WDR70 mutations were identified in ovarian cancer specimens.
Conclusions:
- WDR70 functions as a DNA damage repair gene in vitro.
- WDR70 disruption impairs homologous recombination and causes chromosomal instability.
- Frequent WDR70 mutations in ovarian cancer suggest a role in DNA repairdefects and potential anti-cancer activity.
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