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Published on: August 23, 2016
SLX4 and XPF are involved in cell migration and EMT in a cell-specific manner
Emeline Cros-Perrial1, Sabine Beaumel1, Manon Gimbert1
1Université Claude Bernard Lyon 1, INSERM U-1052, CNRS 5286, Centre Léon Bérard, Centre de Recherche en Cancérologie de Lyon 69008 Lyon, France.
Abstract:
SLX4 and XPF are two proteins involved in DNA repair, but very little is known about their potential roles in other processes of cancer cell biology. We developed original cell models with CRISPR-Cas9-mediated knock-out of SLX4 and/or XPF using five different cell lines (A549, NCI-H1703, COLO-357, HT-29 and HEK-293 T), and performed characterization with cell biology experiments including migration assays, drug sensitivity testing, cell proliferation assessment and Western blots for relevant proteins. Results showed decreased migration of all models in HT-29 cells, of XPF-deficient COLO-357 cells and of SLX4-deficient HEK-293 T cells. Modified cell models had overall increased sensitivity to cisplatin and mitomycine C, and some models showed an increased frequency of double-stranded DNA damages. One NCI-H1703 cell model showed major karyotypic modifications, and epithelial to mesenchymal transition (EMT)-related proteins were modified in several models. Finally, knocking out one or both proteins in A549 cells had not the same impact on in vivo growth in mice. These original cell models allowed us to identify new and DNA repair-unrelated cellular roles of SLX4 and XPF in cancer cell biology. Our results should be considered within work on Nucleotide Excision Repair (NER) inhibition targeting SLX, XPF or other related proteins.
Insights
This study reveals novel, DNA repair-independent functions of SLX4 and XPF proteins in cancer. These findings are crucial for developing new cancer therapies targeting Nucleotide Excision Repair (NER) pathways.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- SLX4 and XPF proteins are known for their roles in DNA repair.
- Their functions beyond DNA repair in cancer cell biology remain largely unexplored.
Purpose of the Study:
- To investigate the non-DNA repair roles of SLX4 and XPF in cancer cell biology.
- To develop and characterize novel cancer cell models with SLX4 and/or XPF knock-out.
Main Methods:
- CRISPR-Cas9 gene editing was used to create SLX4 and/or XPF knock-out cell models in five different human cancer cell lines.
- Characterization involved migration assays, drug sensitivity testing (cisplatin, mitomycine C), proliferation assessment, Western blotting, and karyotyping.
Main Results:
- Knock-out models exhibited decreased cell migration in specific cell lines (HT-29, COLO-357, HEK-293T).
- Increased sensitivity to cisplatin and mitomycine C was observed across modified models.
- Some models showed increased double-stranded DNA damage, karyotypic modifications, and altered epithelial to mesenchymal transition (EMT)-related proteins.
- In vivo growth in mice differed between SLX4/XPF knock-out A549 cell models.
Conclusions:
- SLX4 and XPF possess significant roles in cancer cell biology independent of their DNA repair functions.
- These findings provide new insights for targeting SLX, XPF, or related proteins in Nucleotide Excision Repair (NER) inhibition strategies for cancer treatment.
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