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Targeting Chk2 improves gastric cancer chemotherapy by impairing DNA damage repair

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Abstract

Our results demonstrate that the addition of cisplatin after paclitaxel-induced mitotic arrest was more effective than individual treatment on gastric adenocarcinoma cells (MKN45). However, the treatment did not induce benefits in cells derived from lymph node metastasis (ST2957). Time-lapse microscopy revealed that cell death was caused by mitotic catastrophe and apoptosis induction, as the use of the caspase inhibitor z-VAD-fmk decreased cell death. We propose that the molecular mechanism mediating this cell fate is a slippage suffered by these cells, given that our Western blot (WB) analysis revealed premature cyclin B degradation. This resulted in the cell exiting from mitosis without undergoing DNA damage repair, as demonstrated by the strong phosphorylation of H2AX. A comet assay indicated that DNA repair was impaired, and Western blotting showed that the Chk2 protein was degraded after sequential treatment (paclitaxel-cisplatin). Based on these results, the modulation of cell death during mitosis may be an effective strategy for gastric cancer therapy.

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Abbreviations

CDDP:

Cisplatin

PXL:

Paclitaxel

SAC:

Spindle assembly checkpoint

DDR:

DNA damage response

DSBs:

Double strand breaks

ATM:

Ataxia-telangiectasia mutated

ATR:

ATM- and Rad3-related

γH2AX:

Phosphorylated H2AX

CIN:

Chromosome instability

BRCA1:

Breast cancer 1

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Acknowledgments

We are grateful to Javier Perez, Daniel Gomez (photography facility), Diego Navarro and Lucia Sanchez (microscopy facility IIBM) and Diego Mejias (microscopy facility from CNIO) for technical assistance. We also would like to thank Dr. Marcos Malumbres for the GFP-H4B plasmid, R. Sanchez and Marta Fernandez-Fuente for proofreading the manuscript. This work was supported by the following Grants: PS09/1988, PI11-00949 and CCG10-UAM/BIO-5871. The authors declare no competing relationship or commercial affiliations or financial interests.

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Authors and Affiliations

  1. Department of Biochemistry, School of Medicine, UAM, Madrid, Spain

    A. Gutiérrez-González, J. Bargiela-Iparraguirre & I. Sanchez-Perez

  2. Biomedical Research Institute of Madrid, Madrid CSIC/UAM, C/Arturo Duperier 4, 28029, Madrid, Spain

    A. Gutiérrez-González, J. Bargiela-Iparraguirre, R. Perona & I. Sanchez-Perez

  3. Thoracic, Head & Neck and Brain Oncology Unit, CIOCC Centro integral Oncológico Clara Campal. GHM Grupo Hospital de Madrid, Madrid, Spain

    C. Belda-Iniesta

  4. Biomarkers and Experimental Therapeutics Group, IdiPAZ, University Hospital La Paz, Madrid, Spain

    C. Belda-Iniesta & R. Perona

  5. Department of Medical Oncology, University Hospital Puerta de Hierro, UAM, Madrid, Spain

    G. Dominguez

  6. Department of Medical Oncology, Gregorio Marañon Hospital, Madrid, Spain

    P. García Alfonso

  7. CIBER on Rare Diseases (CIBERER), Valencia, Spain

    R. Perona

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  1. A. Gutiérrez-González
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  2. C. Belda-Iniesta
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  7. I. Sanchez-Perez
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Correspondence to I. Sanchez-Perez.

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Gutiérrez-González, A., Belda-Iniesta, C., Bargiela-Iparraguirre, J. et al. Targeting Chk2 improves gastric cancer chemotherapy by impairing DNA damage repair. Apoptosis 18, 347–360 (2013). https://doi.org/10.1007/s10495-012-0794-2

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  • DOI: https://doi.org/10.1007/s10495-012-0794-2

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