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RGC32, a novel p53-inducible gene, is located on centrosomes during mitosis and results in G2/M arrest

Oncogene volume 26, pages 1110–1121 (2007)Cite this article

Abstract

To identify target genes for the hemizygous deletions of chromosome 13 that are recurrently observed in malignant gliomas, we performed genome-wide DNA copy-number analysis using array-based comparative genomic hybridization and gene expression analysis using an oligonucleotide-array. The response gene to complement 32 (RGC32) at 13q14.11 was identified as a deletion target, and its expression was frequently silenced in glioma cell lines compared with normal brain. Levels of RGC32 mRNA tended to decrease toward higher grades of primary astrocytomas, especially in tumors with mutations of p53. Expression of RGC32 mRNA was dramatically increased by exogenous p53 in a p53-mutant glioma cell line, and also by endogenous p53 in response to DNA damage in p53+/+ colon-cancer cells, but not in isogenic p53−/− cells. Chromatin immunoprecipitation and reporter assays demonstrated binding of endogenous p53 protein to the promoter region of the RGC32 gene, implying p53-dependent transcriptional activity. Transiently and stably overexpressed RGC32 suppressed the growth of glioma cells, probably owing to induction of G2/M arrest. Immunocytochemical analysis revealed a concentration of RGC32 protein at the centrosome during mitosis. RGC32 formed a protein complex with polo-like kinase 1 and was phosphorylated in vitro. These observations implied a novel mechanism by which p53 might negatively regulate cell-cycle progression by way of this newly identified transcriptional target. Our results provide the first evidence that RGC32 might be a possible tumor-suppressor for glioma, that it is directly induced by p53, and that it mediates the arrest of mitotic progression.

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References

  • Badea T, Niculescu F, Soane L, Fosbrink M, Sorana H, Rus V et al. (2002). RGC-32 increases p34CDC2 kinase activity and entry of aortic smooth muscle cells into S-phase. J Biol Chem 277: 502–508.

    Article  CAS  Google Scholar 

  • Badea TC, Niculescu FI, Soane L, Shin ML, Rus H . (1998). Molecular cloning and characterization of RGC-32, a novel gene induced by complement activation in oligodendrocytes. J Biol Chem 273: 26977–26981.

    Article  CAS  Google Scholar 

  • Barr FA, Sillje HH, Nigg EA . (2004). Polo-like kinases and the orchestration of cell division. Nat Rev Mol Cell Biol 5: 429–440.

    Article  CAS  Google Scholar 

  • Burns KL, Ueki K, Jhung SL, Koh J, Louis DN . (1998). Molecular genetic correlates of p16, cdk4, and pRb immunohistochemistry in glioblastomas. J Neuropathol Exp Neurol 57: 122–130.

    Article  CAS  Google Scholar 

  • Dietzmann K, Kirches E, von Bossanyi P, Jachau K, Mawrin C . (2001). Increased human polo-like kinase-1 expression in gliomas. J Neurooncol 53: 1–11.

    Article  CAS  Google Scholar 

  • el-Deiry WS, Kern SE, Pietenpol JA, Kinzler KW, Vogelstein B . (1992). Definition of a consensus binding site for p53. Nat Genet 1: 45–49.

    Article  CAS  Google Scholar 

  • Elez R, Piiper A, Giannini CD, Brendel M, Zeuzem S . (2000). Polo-like kinase1, a new target for antisense tumor therapy. Biochem Biophys Res Commun 269: 352–356.

    Article  CAS  Google Scholar 

  • Gumireddy K, Reddy MV, Cosenza SC, Boominathan R, Baker SJ, Papathi N et al. (2005). ON01910, a non-ATP-competitive small molecule inhibitor of Plk1, is a potent anticancer agent. Cancer Cell 7: 275–286.

    Article  CAS  Google Scholar 

  • Holland EC . (2001). Gliomagenesis: genetic alterations and mouse models. Nat Rev Genet 12: 120–129.

    Article  Google Scholar 

  • Imoto I, Yuki Y, Sonoda I, Ito T, Shimada Y, Imamura M et al. (2003). Identification of ZASC1 encoding a Kruppel-like zinc finger protein as a novel target for 3q26 amplification in esophageal squamous cell carcinomas. Cancer Res 63: 5691–5696.

    CAS  Google Scholar 

  • Inazawa J, Inoue J, Imoto I . (2004). Comparative genomic hybridization (CGH)-arrays pave the way for identification of novel cancer-related genes. Cancer Sci 95: 559–563.

    Article  CAS  Google Scholar 

  • Inda MM, Fan X, Munoz J, Perot C, Fauvet D, Danglot G et al. (2003). Chromosomal abnormalities in human glioblastomas: gain in chromosome 7p correlating with loss in chromosome 10q. Mol Carcinog 36: 6–14.

    Article  Google Scholar 

  • Inoue J, Otsuki T, Hirasawa A, Imoto I, Matsuo Y, Shimizu S et al. (2004). Overexpression of PDZK1 within the 1q12–q22 amplicon is likely to be associated with drug-resistance phenotype in multiple myeloma. Am J Pathol 165: 71–81.

    Article  CAS  Google Scholar 

  • Jongsma AP, Piek JM, Zweemer RP, Verheijen RH, Klein Gebbinck JW, van Kamp GJ et al. (2002). Molecular evidence for putative tumour suppressor genes on chromosome 13q specific to BRCA1 related ovarian and fallopian tube cancer. Mol Pathol 55: 305–309.

    Article  CAS  Google Scholar 

  • Kleihues P, Louis DN, Scheithauer BW, Rorke LB, Reifenberger G, Burger PC et al. (2002). The WHO classification of tumors of the nervous system. J Neuropathol Exp Neurol 61: 215–225.

    Article  Google Scholar 

  • Ko LJ, Prives C . (1996). p53: puzzle and paradigm. Genes Dev 10: 1054–1072.

    Article  CAS  Google Scholar 

  • Kunwar S, Mohapatra G, Bollen A, Lamborn KR, Prados M, Feuerstein BG . (2001). Genetic subgroups of anaplastic astrocytomas correlate with patient age and survival. Cancer Res 61: 7683–7688.

    CAS  PubMed  Google Scholar 

  • Lane HA, Nigg EA . (1996). Antibody microinjection reveals an essential role for human polo-like kinase 1 (Plk1) in the functional maturation of mitotic centrosomes. J Cell Biol 135: 1701–1713.

    Article  CAS  Google Scholar 

  • Liu X, Erikson RL . (2002). Activation of Cdc2/cyclin B and inhibition of centrosome amplification in cells depleted of Plk1 by siRNA. Proc Natl Acad Sci USA 99: 8672–8676.

    Article  CAS  Google Scholar 

  • Liu X, Erikson RL . (2003). Polo-like kinase (Plk)1 depletion induces apoptosis in cancer cells. Proc Natl Acad Sci USA 100: 5789–5794.

    Article  CAS  Google Scholar 

  • Lu W, Takahashi H, Furusato M, Maekawa S, Nakano M, Meng C et al. (2006). Allelotyping analysis at chromosome 13q of high-grade prostatic intraepithelial neoplasia and clinically insignificant and significant prostate cancers. Prostate 66: 405–412.

    Article  CAS  Google Scholar 

  • Maher EA, Furnari FB, Bachoo RM, Rowitch DH, Louis DN, Cavenee WK et al. (2001). Malignant glioma: genetics and biology of a grave matter. Genes Dev 15: 1311–1333.

    Article  CAS  Google Scholar 

  • Marumoto T, Hirota T, Morisaki T, Kunitoku N, Zhang D, Ichikawa Y et al. (2002). Roles of aurora-A kinase in mitotic entry and G2 checkpoint in mammalian cells. Genes Cells 7: 1173–1182.

    Article  CAS  Google Scholar 

  • Misawa A, Inoue J, Sugino Y, Hosoi H, Sugimoto T, Hosoda F et al. (2005). Methylation-associated silencing of the nuclear receptor 1I2 gene in advanced-type neuroblastomas, identified by bacterial artificial chromosome array-based methylated CpG island amplification. Cancer Res 65: 10233–10242.

    Article  CAS  Google Scholar 

  • Miyashita T, Reed JC . (1995). Tumor suppressor p53 is a direct transcriptional activator of the human bax gene. Cell 80: 293–299.

    Article  CAS  Google Scholar 

  • Mundt KE, Golsteyn RM, Lane HA, Nigg EA . (1997). On the regulation and function of human polo-like kinase 1 (PLK1): effects of overexpression on cell cycle progression. Biochem Biophys Res Commun 239: 377–385.

    Article  CAS  Google Scholar 

  • Nakamura M, Watanabe T, Yonekawa Y, Kleihues P, Ohgaki H . (2001). Promoter methylation of the DNA repair gene MGMT in astrocytomas is frequently associated with G:C → A:T mutations of the TP53 tumor suppressor gene. Carcinogenesis 22: 1715–1719.

    Article  CAS  Google Scholar 

  • Nishizaki T, Ozaki S, Harada K, Ito H, Arai H, Beppu T et al. (1998). Investigation of genetic alterations associated with the grade of astrocytic tumor by comparative genomic hybridization. Genes Chromosomes Cancer 21: 340–346.

    Article  CAS  Google Scholar 

  • Oda K, Arakawa H, Tanaka T, Matsuda K, Tanikawa C, Mori T et al. (2000). p53AIP1, a potential mediator of p53-dependent apoptosis, and its regulation by Ser-46-phosphorylated p53. Cell 102: 849–862.

    Article  CAS  Google Scholar 

  • Ohgaki H, Dessen P, Jourde B, Horstmann S, Nishikawa T, Di Patre PL et al. (2004). Genetic pathways to glioblastoma: a population-based study. Cancer Res 64: 6892–6899.

    Article  CAS  Google Scholar 

  • Okamoto Y, Di Patre PL, Burkhard C, Horstmann S, Jourde B, Fahey M et al. (2004). Population-based study on incidence, survival rates, and genetic alterations of low-grade diffuse astrocytomas and oligodendrogliomas. Acta Neuropathol (Berlin) 108: 49–56.

    Article  Google Scholar 

  • Pan H, Califano J, Ponte JF, Russo AL, Cheng KH, Thiagalingam A et al. (2005). Loss of heterozygosity patterns provide fingerprints for genetic heterogeneity in multistep cancer progression of tobacco smoke-induced non-small cell lung cancer. Cancer Res 65: 1664–1669.

    Article  CAS  Google Scholar 

  • Saigusa K, Hashimoto N, Tsuda H, Yokoi S, Maruno M, Yoshimine T et al. (2005). Overexpressed Skp2 within 5p amplification detected by array-based comparative genomic hybridization is associated with poor prognosis of glioblastomas. Cancer Sci 96: 676–683.

    Article  CAS  Google Scholar 

  • Sato T, Akiyama F, Sakamoto G, Kasumi F, Nakamura Y . (1991). Accumulation of genetic alterations and progression of primary breast cancer. Cancer Res 51: 5794–5799.

    CAS  Google Scholar 

  • Shaughnessy J, Tian E, Sawyer J, Bumm K, Landes R, Badros A et al. (2000). High incidence of chromosome 13 deletion in multiple myeloma detected by multiprobe interphase FISH. Blood 96: 1505–1511.

    CAS  Google Scholar 

  • Sonoda I, Imoto I, Inoue J, Shibata T, Shimada Y, Chin K et al. (2004). Frequent silencing of low density lipoprotein receptor-related protein 1B (LRP1B) expression by genetic and epigenetic mechanisms in esophageal squamous cell carcinoma. Cancer Res 64: 3741–3747.

    Article  CAS  Google Scholar 

  • Spankuch-Schmitt B, Bereiter-Hahn J, Kaufmann M, Strebhardt K . (2002). Effect of RNA silencing of polo-like kinase-1 (PLK1) on apoptosis and spindle formation in human cancer cells. J Natl Cancer Inst 94: 1863–1877.

    Article  CAS  Google Scholar 

  • Takada H, Imoto I, Tsuda H, Sonoda I, Ichikura T, Mochizuki H et al. (2005). Screening of DNA copy-number aberrations in gastric cancer cell lines by array-based comparative genomic hybridization. Cancer Sci 96: 100–110.

    Article  CAS  Google Scholar 

  • Takai N, Hamanaka R, Yoshimatsu J, Miyakawa I . (2005). Polo-like kinases (Plks) and cancer. Oncogene 24: 287–291.

    Article  CAS  Google Scholar 

  • Tokino T, Thiagalingam S, el-Deiry WS, Waldman T, Kinzler KW, Vogelstein B . (1994). p53 tagged sites from human genomic DNA. Hum Mol Genet 3: 1537–1542.

    Article  CAS  Google Scholar 

  • Zhu Y, Parada LF . (2002). The molecular and genetic basis of neurological tumours. Nat Rev Cancer 2: 616–626.

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We thank Dr Bert Vogelstein (Howard Hughes Medical Institute, The Sidney Kimmel Comprehensive Cancer Center, and Program in Cellular and Molecular Medicine, The Johns Hopkins University Medical Institutions) for HCT116 (p53+/+) and HCT116 (p53−/−) cell lines and Dr Hideyuki Saya (Department of Tumor Genetics and Biology, Graduate School of Medical Sciences, Kumamoto University) for the anti-Aurora-A antibody. We also thank Ai Watanabe, Ayako Takahashi and Rumi Mori for technical assistance. This work was supported by Grants-in-Aid for Scientific Research on Priority Areas (C) from the Ministry of Education, Culture, Sports, Science, and Technology, Japan; by a Grant-in-Aid from Core Research for Evolutional Science and Technology (CREST) of the Japan Science and Technology Corporation (JST); and by 21st Century Center of Excellence Program for Molecular Destruction and Reconstitution of Tooth and Bone.

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Authors and Affiliations

  1. Department of Molecular Cytogenetics, Medical Research Institute and Graduate School of Biomedical Science, Tokyo Medical and Dental University, Tokyo, Japan

    K Saigusa, I Imoto & J Inazawa

  2. Department of Neurosurgery, Graduate School, Tokyo Medical and Dental University, Tokyo, Japan

    K Saigusa, M Aoyagi & K Ohno

  3. Core Research for Evolutional Science and Technology (CREST) of Japan Science and Technology Corporation (JST), Saitama, Japan

    K Saigusa, I Imoto & J Inazawa

  4. Hard Tissue Genome research Center, Tokyo Medical and Dental University, Tokyo, Japan

    I Imoto & J Inazawa

  5. Human Genome Center, The Institute of Medical Science, University of Tokyo, Tokyo, Japan

    C Tanikawa & Y Nakamura

  6. 21st Century Center of Excellence Program for Molecular Destruction and Reconstitution of Tooth and Bone, Tokyo Medical and Dental University, Tokyo, Japan

    J Inazawa

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  1. K Saigusa
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Correspondence to J Inazawa.

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Saigusa, K., Imoto, I., Tanikawa, C. et al. RGC32, a novel p53-inducible gene, is located on centrosomes during mitosis and results in G2/M arrest. Oncogene 26, 1110–1121 (2007). https://doi.org/10.1038/sj.onc.1210148

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