Abstract
Propose
Poly (ADP-ribose) polymerase 1 inhibitors were originally investigated as anti-cancer therapeutics with BRCA1/2 genes mutation. Here, we investigate the effectiveness of a novel PARP1 inhibitor fluzoparib, for enhancing the radiation sensitivity of NSCLC cells lacking BRCA1/2 mutation.
Methods
We used MTS assays, western blotting, colony formation assays, immunofluorescence staining, and flow cytometry to evaluate the radiosensitization of NSCLC cells to fluzoparib and explore the underlying mechanisms in vitro. Through BRCA1 and RAD50 genes knockdown, we established dysfunctional homologous recombination (HR) DNA repair pathway models in NSCLC cells. We next investigated the radiosensitization effect of fluzoparib in vivo using human NSCLC xenograft models in mice. The expression of PARP1 and BRCA1 in human NSCLC tumor samples was measured by immunohistochemistry. Furthermore, we sequenced HR-related gene mutations and analyzed their frequencies in advanced NSCLC.
Results
In vitro experiments in NSCLC cell lines along with in vivo experiments using an NSCLC xenograft mouse model demonstrated the radiosensitization effect of fluzoparib. The underlying mechanisms involved increased apoptosis, cell-cycle arrest, enhanced irradiation-induced DNA damage, and delayed DNA-damage repair. Immunohistochemical staining showed no correlation between the expression of PARP1 and BRCA1. Moreover, our sequencing results revealed high mutation frequencies for the BRCA1/2, CHEK2, ATR, and RAD50 genes.
Conclusion
The potential therapeutic value of fluzoparib for increasing the radiation sensitivity of NSCLC is well confirmed. Moreover, our findings of high mutation frequencies among HR genes suggest that PARP1 inhibition may be an effective treatment strategy for advanced non-small cell lung cancer patients.
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Acknowledgements
We wish to thank the timely help given by Dr. Qinghua Wang and Dr. Shuai Ma in data analysis and plotting.
Funding
This research was supported by a Grant from Tianjin Natural Science Foundation (16jcybjc25300).
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Conception and design: NBL, GGY, LJZ, and PW. Development of methodology: NBL, GGY, LJZ, and CYY. Acquisition of data (provided animals, acquired and managed patients, provided facilities, etc.): JL, HH, XCD, LMZ, JC, JFS, QW, YX, and SLX. Analysis and interpretation of data (e.g., statistical analysis, biostatistics, and computational analysis): JL, HH, XCD, and NBL. Writing, review, and/or revision of the manuscript: JL and NBL. Administrative, technical, or material support (i.e., reporting or organizing data, constructing databases): YX, SLX, and GGY. Study supervision: NBL and GGY.
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Luo, J., Dai, X., Hu, H. et al. Fluzoparib increases radiation sensitivity of non-small cell lung cancer (NSCLC) cells without BRCA1/2 mutation, a novel PARP1 inhibitor undergoing clinical trials. J Cancer Res Clin Oncol 146, 721–737 (2020). https://doi.org/10.1007/s00432-019-03097-6
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DOI: https://doi.org/10.1007/s00432-019-03097-6