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The checkpoint 1 kinase inhibitor LY2603618 induces cell cycle arrest, DNA damage response and autophagy in cancer cells

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Abstract

Chemotherapy- or radiotherapy-induced DNA damage activates the Chk1-dependent DNA damage response (DDR) and cell cycle checkpoints to facilitate cell survival. Numerous attempts have been made to identify specific Chk1 inhibitors to enhance the efficiency of chemotherapy or radiotherapy. In this study, we investigated the molecular mechanisms underlying the antitumor activity of LY2603618, a potent and selective small molecule inhibitor of Chk1 protein kinase, in human lung cancer cells. Treatment of cancer cells with LY2603618 caused cell cycle arrest in the G2/M phase. A marked induction of DDR, including the phosphorylation of ATM, Chk2, p53 and histone H2AX, was observed after LY2603618 treatment. LY2603618 inhibited Chk1 autophosphorylation (S296 Chk1) and increased DNA damage-mediated Chk1 phosphorylation (S345 Chk1). In addition, LY2603618-treated lung cancer cells transitioned from LC3-I to LC3-II, a hallmark of autophagy. Blocking autophagy with chloroquine (CQ) further enhanced LY2603618′s inhibitory effect on cell viability/proliferation. LY2603618 also significantly increased p38 and c-Jun N-terminal kinase (JNK) phosphorylation. Pretreatment with the JNK inhibitor reduced cleavage of caspase-3 and PARP levels in LY2603618-treated cells. These results suggest the following: (i) the biological consequences of LY2603618 in lung cancer cells is associated with both inhibition of Chk1 phosphorylation on S296 and activation of the DNA damage response network; and (ii) the anticancer property of LY2603618 might be increased by inhibiting autophagy.

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 81272683 to Feng-Ze Wang, No. 81271275 to Bao-Liang Sun).

Conflict of interest

The authors declare that there are no conflicts of interest.

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Authors and Affiliations

  1. School of Biological Science, Taishan Medical University, Taian, 271016, People’s Republic of China

    Feng-Ze Wang, Zhao-Mei Li & Xue-Ying Wang

  2. Key Lab of cerebral microcirculation in Universities of Shandong (Taishan Medical University), Taishan Medical University, Taian, 271000, People’s Republic of China

    Feng-Ze Wang, Xiao-Yi Yang & Bao-Liang Sun

  3. School of Pharmacology, Taishan Medical University, Taian, 271016, People’s Republic of China

    Hong-rong Fei & Ji-Guo Zhang

  4. Institute of Atherosclerosis, Taishan Medical University, Taian, 271000, People’s Republic of China

    Ying-Jie Cui

  5. College of Landscape Architecture and Forestry, Qingdao Agricultural University, Qingdao, 266109, People’s Republic of China

    Ying-Kun Sun

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  1. Feng-Ze Wang
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  2. Hong-rong Fei
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Correspondence to Ji-Guo Zhang or Bao-Liang Sun.

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Feng-Ze Wang and Hong-rong Fei have contributed equally to this work.

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Wang, FZ., Fei, Hr., Cui, YJ. et al. The checkpoint 1 kinase inhibitor LY2603618 induces cell cycle arrest, DNA damage response and autophagy in cancer cells. Apoptosis 19, 1389–1398 (2014). https://doi.org/10.1007/s10495-014-1010-3

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