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The newly synthesized 2-arylnaphthyridin-4-one, CSC-3436, induces apoptosis of non-small cell lung cancer cells by inhibiting tubulin dynamics and activating CDK1

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Abstract

Purpose

To investigate the anticancer therapeutic potential of a new synthetic compound, 2-(3-hydroxyphenyl)-5-methylnaphthyridin-4-one (CSC-3436), on non-small cell lung cancer (NSCLC) cells.

Methods

Cell viability was determined by MTT assay. Cell cycle distribution was assessed by propidium iodide staining and subjected to flow cytometry analysis. Protein expression was detected by western blot analysis. Pharmacological inhibitors and shRNAs were applied to examine the possible pathways involved CSC-3436-inhibited viability of NSCLC cells.

Results

CSC-3436 decreased NSCLC cell viability by inducing apoptosis. In vivo and in vitro tubulin polymerization assays revealed that CSC-3463 caused tubulin depolymerization by directly binding to the colchicine-binding site. Furthermore, CSC-3436 caused the mitotic arrest with a marked activation of cyclin-dependent kinase 1 (CDK1) and increased the expression of phospho-Ser/Thr-Pro mitotic protein monoclonal 2. The CDK1 inhibitor, roscovitine, reversed the CSC-3436-induced upregulation of CDK1 activity as well as the mitotic arrest. DNA damage response kinases, including ataxia telangiectasia mutated (ATM), ATM and Rad3-related, DNA-dependent protein kinase, checkpoint kinase 1, and checkpoint kinase 2, were phosphorylated and activated by CSC-3436. c-Jun N-terminal kinase was activated by CSC-3436 and involved in the regulation of mitotic arrest and apoptosis. CSC-3436-induced apoptosis was accompanied by the activation of pro-apoptotic factors FADD, TRADD, and RIP and the inactivation of anti-apoptotic proteins Bcl-2 and Bcl-xL, resulting in the cleavage and subsequent activation of caspases.

Conclusions

Our results reveal the cellular events in which CSC-3436 induces tumor cell death and demonstrate that CSC-3436 is a potential tubulin-disrupting agent for antitumor therapy against NSCLC.

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Abbreviations

ATM:

Ataxia telangiectasia mutated

ATR:

ATM and Rad3-related

CDK1:

Cyclin-dependent kinase 1

Chk1:

Checkpoint kinase 1

Chk2:

Checkpoint kinase 2

CSC-3436:

2-(3-Hydroxyphenyl)-5-methylnaphthyridin-4-one

DAPI:

4ʹ,6ʹ-Diamino-2-phenylindole

DNA-PK:

DNA-dependent protein kinase

FBS:

Fetal bovine serum

FITC:

Fluorescein isothiocyanate

JNK:

c-Jun N-terminal kinase

MPM-2:

Phospho-Ser/Thr-Pro mitotic protein monoclonal 2

NSCLC:

Non-small cell lung cancer

PARP:

Poly(ADP-ribose)polymerase

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Acknowledgments

This work was supported by the National Science Council of the Republic of China (Grant Numbers NSC102-2325-B-039-005 and NSC102-2320-B-039-012 to S. C. Kuo, and NSC102-2321-B-039-004 to Y. L. Yu), the National Health Research Institute of the Republic of China (Grant Number NHRI-EX102-10245BI to Y. L. Yu), China Medical University Hospital (Grant Number DMR-104-097 and DMR-104-108 to L.-C. Chang), and CMU under the Aim for Top University Plan of the Ministry of Education, Taiwan.

Conflict of interest

No potential conflicts of interest were disclosed.

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    Authors and Affiliations

    1. Chinese Medicinal Research and Development Center, China Medical University Hospital, No. 2, Yude Road, Taichung, 404, Taiwan, ROC

      Ling-Chu Chang, Min-Tsang Hsieh & Yang-Chang Wu

    2. Graduate Institute of Cancer Biology, Center for Molecular Medicine, China Medical University, Taichung, 404, Taiwan, ROC

      Yung-Luen Yu, Ruey-Hwang Chou, Yang-Chang Wu & Sheng-Chu Kuo

    3. Department of Biotechnology, Asia University, Taichung, 413, Taiwan

      Yung-Luen Yu & Ruey-Hwang Chou

    4. Graduate Institute of Pharmaceutical Chemistry, China Medical University, No. 91, Hsueh-Shih Road, Taichung, 404, Taiwan, ROC

      Chin-Yu Liu, Yung-Yi Cheng, Hui-Yi Lin & Li-Jiau Huang

    5. School of Pharmacy, China Medical University, Taichung, 404, Taiwan, ROC

      Min-Tsang Hsieh & Yang-Chang Wu

    6. Institute of Clinical Pharmacy and Pharmaceutical Sciences, National Cheng Kung University, Tainan, 701, Taiwan, ROC

      Hsin-Yi Hung

    7. Graduate Institute of Natural Products, Kaohsiung Medical University, Kaohsiung, Taiwan, ROC

      Yang-Chang Wu

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    Correspondence to Ling-Chu Chang or Sheng-Chu Kuo.

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    Ling-Chu Chang and Yung-Luen Yu have contributed equally to this work.

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    Chang, LC., Yu, YL., Liu, CY. et al. The newly synthesized 2-arylnaphthyridin-4-one, CSC-3436, induces apoptosis of non-small cell lung cancer cells by inhibiting tubulin dynamics and activating CDK1. Cancer Chemother Pharmacol 75, 1303–1315 (2015). https://doi.org/10.1007/s00280-015-2765-0

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