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DFF45/ICAD restores cisplatin-induced nuclear fragmentation but not DNA cleavage in DFF45-deficient neuroblastoma cells

Oncogene volume 26, pages 5669–5673 (2007)Cite this article

Abstract

We have previously defined a homozygously deleted region at chromosome 1p36.2–p36.3 in human neuroblastoma cell lines, NB-1 and NB-C201, and identified six genes including DFF45/ICAD within this region. In this study, we found that NB-C201 cells are much more resistant to various genotoxic stresses such as cisplatin (CDDP) than CHP134 and SH-SY5Y cells that do not have the homozygous deletion. To examine a role(s) of DFF45 in the regulation of apoptosis in response to CDDP, we have established stably DFF45-expressing NB-C201 cell clones (DFF45-1 and DFF45-3) and a control cell clone (NB-C201-C) using a retrovirus-mediated gene transfer. In contrast to NB-C201-C cells, DFF45-3 cells displayed apoptotic nuclear fragmentation in response to CDDP. Although CDDP-induced proteolytic cleavage of procaspase-3 and DFF45 in DFF45-3 cells, we could not detect a typical apoptotic DNA fragmentation. Additionally, deletion analysis revealed that C-terminal region of DFF45 is required for inducing nuclear fragmentation. Unexpectedly, (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays demonstrated that DFF45 has undetectable effect on CDDP sensitivity of NB-C201 cells. Taken together, our present results suggest that DFF45/DFF40 system may be sufficient for CDDP-induced nuclear fragmentation but not DNA cleavage.

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References

  • Amler LC, Corvi R, Praml C, Savelyeva L, Le Paslier D, Schwab M . (1995). A reciprocal translocation (1;15)(36.2;q24) in a neuroblastoma cell line is accompanied by DNA duplication and may signal the site of a putative tumor suppressor gene. Oncogene 10: 1095–1101.

    CAS  PubMed  Google Scholar 

  • Brodeur GM, Seeger RC, Schwab M, Varmas HE, Bishop JM . (1984). Amplification of N-myc in untreated human neuroblastomas corelates with advanced disease stage. Science 224: 1121–1124.

    Article  CAS  Google Scholar 

  • Brodeur GM, Nakagawara A . (1992). Molecular basis of clinical heterogeneity in neuroblastoma. Am J Pediatr Hematol Oncol 14: 111–116.

    Article  CAS  Google Scholar 

  • Cheng NC, Van Roy N, Chan A, Beitsma M, Westerveld A, Speleman F et al. (1995). Deletion mapping in neuroblastoma cell lines suggests two distinct tumor suppressor genes in the 1p35–36 region, only one of which is associated with N-myc amplification. Oncogene 10: 291–297.

    CAS  PubMed  Google Scholar 

  • Enari M, Sakahira H, Yokoyama H, Okawa K, Iwamatsu A, Nagata S . (1998). A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391: 43–50.

    Article  CAS  Google Scholar 

  • Gu J, Dong RP, Zhang C, McLaughlin DF, Wu MX, Schlossman SF . (1999). Functional interaction of DFF35 and DFF45 with caspase-activated DNA fragmentation nuclease DFF40. J Biol Chem 274: 20759–20762.

    Article  CAS  Google Scholar 

  • Halenbeck R, MacDonald H, Roulston A, Chen TT, Conroy L, Williams LT . (1998). CPAN, a human nuclease regulated by the caspase-sensitive inhibitor DFF45. Curr Biol 8: 537–540.

    Article  CAS  Google Scholar 

  • Hosoda M, Ozaki T, Miyazaki K, Hayashi S, Furuya S, Watanabe K et al. (2005). UFD2a mediates the proteasomal turnover of p73 without promoting p73 ubiquitination. Oncogene 24: 7156–7169.

    Article  CAS  Google Scholar 

  • Inohara N, Koseki T, Chen S, Benedict MA, Nunez G . (1999). Identification of regulatory and catalytic domains in the apoptosis nuclease DFF40/CAD. J Biol Chem 274: 270–274.

    Article  CAS  Google Scholar 

  • Liu X, Zou H, Slaughter C, Wang X . (1997). DFF, a heterodimeric protein that functions downstream of caspase-3 to trigger DNA fragmentation during apoptosis. Cell 89: 175–184.

    Article  CAS  Google Scholar 

  • Liu X, Li P, Widlak P, Zou H, Luo X, Garrard WT et al. (1998). The 40-kDa subunit of DNA fragmentation factor induces DNA fragmentation and chromatin condensation during apoptosis. Proc Natl Acad Sci USA 95: 8461–8466.

    Article  CAS  Google Scholar 

  • Lu C, Zhu F, Cho YY, Tang F, Zykova T, Ma WY et al. (2006). Cell apoptosis: requirement of H2AX in DNA ladder formation, but not for the activation of caspase-3. Mol Cell 23: 121–132.

    Article  CAS  Google Scholar 

  • Lu Z, Zhang C, Zhai Z . (2005). Nucleoplasmin regulates chromatin condensation during apoptosis. Proc Natl Acad Sci USA 102: 2778–2783.

    Article  CAS  Google Scholar 

  • Martinsson T, Sjoberg RM, Hedborg F, Kogner P . (1995). Deletion of chromosome 1p loci and microsatellite instability in neuroblastomas analyzed with short-tandem repeat polymorphisms. Cancer Res 55: 5681–5686.

    CAS  PubMed  Google Scholar 

  • Mukae N, Enari M, Sakahira H, Fukuda Y, Inazawa J, Tho H et al. (1998). Molecular cloning and characterization of human caspase- activated DNase. Proc Natl Acad Sci USA 95: 9123–9128.

    Article  CAS  Google Scholar 

  • Ohira M, Kageyama H, Mihara M, Furuta S, Machida T, Shishikura T et al. (2000). Identification and characterization of a 500-kb homozygously deleted region at 1p36.2–p36.3 in a neuroblastoma cell line. Oncogene 19: 4302–4307.

    Article  CAS  Google Scholar 

  • Raff MC . (1992). Social controls on cell survival and cell death. Nature 356: 397–400.

    Article  CAS  Google Scholar 

  • Sakahira H, Enari M, Nagata S . (1998). Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature 391: 96–99.

    Article  CAS  Google Scholar 

  • Schleiermacher G, Peter M, Michon J, Hugot JP, Vielh P, Zucker JM et al. (1994). Two distinct deleted regions on the short arm of chromosome 1 in neuroblastoma. Genes Chromosomes Cancer 10: 275–281.

    Article  CAS  Google Scholar 

  • Takeda O, Homma C, Maseki N, Sakurai M, Kanda N, Schwab M et al. (1994). There may be two tumor suppressor genes on chromosome arm 1p closely associated with biologically distinct subtypes of neuroblastoma. Genes Chromosomes Cancer 10: 30–39.

    Article  CAS  Google Scholar 

  • White PS, Maris JM, Beltinger C, Sulman E, Marshall HN, Fujimori M et al. (1995). A region of consistent deletion in neuroblastoma maps within human chromosome 1p36.2–36.3. Proc Natl Acad Sci USA 92: 5520–5524.

    Article  CAS  Google Scholar 

  • Wyllie AH . (1980). Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284: 555–556.

    Article  CAS  Google Scholar 

  • Yang HW, Chen YZ, Piao HY, Takita J, Soeda E, Hayashi Y . (2001). DNA fragmentation factor 45 (DFF45) gene at 1p36.2 is homozygously deleted and encodes variant transcripts in neuroblastoma cell line. Neoplasia 3: 165–169.

    Article  CAS  Google Scholar 

  • Zhang J, Wang X, Bove KE, Xu M . (1999). DNA fragmentation factor 45-deficient cells are more resistant to apoptosis and exhibit different dying morphology than wild-type control cells. J Biol Chem 274: 37450–37454.

    Article  CAS  Google Scholar 

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Acknowledgements

This work was supported in part by a Grant-in-Aid from the Ministry of Health, Labour and Welfare for Third Term Comprehensive Control Research for Cancer, and a Grant-in-Aid for Cancer Research from the Ministry of Health, Labour and Welfare of Japan, a Grant-in-Aid for Scientific Research on Priority Areas from the Ministry of Education, Culture, Sports, Science and Technology, Japan, and a grant from Uehara Memorial Foundation.

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Authors and Affiliations

  1. Division of Biochemistry, Chiba Cancer Center Research Institute, Chiba, Chuoh-ku, Japan

    M Takahashi, T Ozaki, A Takahashi, N Takada, T Kamijo & A Nakagawara

  2. Department of General Surgery, Hokkaido University School of Medicine, Sapporo, Kita-ku, Japan

    M Takahashi, N Takada & S Todo

  3. Center for Functional Genomics, Hisamitsu Pharmaceutical Co., Inc., Chiba, Chuoh-ku, Japan

    A Takahashi, S Ono & T Koda

  4. Division of Pathology, Chiba Cancer Center Research Institute, Chiba, Chuoh-ku, Japan

    M Miyauchi

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  1. M Takahashi
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Correspondence to A Nakagawara.

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Takahashi, M., Ozaki, T., Takahashi, A. et al. DFF45/ICAD restores cisplatin-induced nuclear fragmentation but not DNA cleavage in DFF45-deficient neuroblastoma cells. Oncogene 26, 5669–5673 (2007). https://doi.org/10.1038/sj.onc.1210352

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