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Co-inhibition of Pol η and ATR sensitizes cisplatin-resistant non-small cell lung cancer cells to cisplatin by impeding DNA damage repair

Acta Pharmacologica Sinicavolume 39pages13591372 (2018) | Download Citation

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Abstract

For the majority of patients with advanced non-small cell lung cancer (NSCLC), the standard of care remains platinum-based chemotherapy. However, cisplatin resistance is a big obstacle to the treatment, and elucidation of its mechanism is warranted. In this study, we showed that there was no difference in intracellular uptake of cisplatin or the removal of platinum-DNA adducts between a cisplatin-resistant NSCLC cell line (A549/DR) and a cisplatin-sensitive NSCLC cell line (A549). However, the capacity to repair DNA interstrand crosslinks (ICLs) and double-strand breaks (DSBs) was significantly enhanced in the A549/DR cell line compared to 3 cisplatin-sensitive cell lines. We found that the protein and mRNA expression levels of Pol η, a Y-family translesion synthesis (TLS) polymerase, were markedly increased upon cisplatin exposure in A549/DR cells compared with A549 cells. Furthermore, intracellular co-localization of Pol η and proliferation cell nuclear antigen (PCNA) induced by cisplatin or cisplatin plus gemcitabine treatment was inhibited by depleting ataxia telangiectasia mutated and Rad-3-related (ATR). Pol η depletion by siRNA sensitized A549/DR cells to cisplatin; co-depletion of Pol η and ATR further increased A549/DR cell death induced by cisplatin or cisplatin plus gemcitabine compared to depletion of Pol η or ATR alone, concomitant with inhibition of DNA ICL and DSB repair and accumulation of DNA damage. No additional sensitization effect of co-depleting Pol η and ATR was observed in A549 cells. These results demonstrate that co-inhibition of Pol η and ATR reverses the drug resistance of cisplatin-resistant NSCLC cells by blocking the repair of DNA ICLs and DSBs induced by cisplatin or cisplatin plus gemcitabine.

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This research was partially supported by the National Youth Science Foundation of China (Grant No 81402485).

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Author notes

  1. These authors contributed equally to this work.

Affiliations

  1. Department of Medical Oncology, Affiliated Hospital of Jiangsu University, Zhenjiang, 212001, China

    • Xiao-qin Li
    • , Jin Ren
    •  & Jian Li
  2. Department of Pulmonary Medicine, Affiliated Hospital of Jiangsu University, Zhenjiang, 212001, China

    • Ping Chen
    • , Yu-jiao Chen
    •  & Kang Chen
  3. Institute of Medical Science, Jiangsu University, Zhenjiang, 212013, China

    • Min Wu
    •  & Yan Wu

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Corresponding author

Correspondence to Jian Li.

Electronic supplementary material

  1. Supplementary Figure S1

    The colony formation of four NSCLC cell lines, and cisplatin-induced the protein and mRNA expressions of TLS polymerases in LOU-NH91 and HCC4006 cell lines.

  2. Supplementary Figure S2

    The viability assay of A549 and A549/DR cell lines after different treatments.

  3. Supplementary Figure S3

    Protein expressions of cleaved caspase-3 and PARP, and DNA replication in A549/DR cells transfected with siRNAs as indicated after drug treatments.

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DOI

https://doi.org/10.1038/aps.2017.187