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Cancer-associated alteration of pericentromeric heterochromatin may contribute to chromosome instability

Oncogene volume 31, pages 3244–3253 (2012)Cite this article

Abstract

Many tumors exhibit elevated chromosome mis-segregation termed chromosome instability (CIN), which is likely to be a potent driver of tumor progression and drug resistance. Causes of CIN are poorly understood but probably include prior genome tetraploidization, centrosome amplification and mitotic checkpoint defects. This study identifies epigenetic alteration of the centromere as a potential contributor to the CIN phenotype. The centromere controls chromosome segregation and consists of higher-order repeat (HOR) alpha-satellite DNA packaged into two chromatin domains: the kinetochore, harboring the centromere-specific H3 variant centromere protein A (CENP-A), and the pericentromeric heterochromatin, considered important for cohesion. Perturbation of centromeric chromatin in model systems causes CIN. As cancer cells exhibit widespread chromatin changes, we hypothesized that pericentromeric chromatin structure could also be affected, contributing to CIN. Cytological and chromatin immunoprecipitation and PCR (ChIP–PCR)-based analyses of HT1080 cancer cells showed that only one of the two HORs on chromosomes 5 and 7 incorporate CENP-A, an organization conserved in all normal and cancer-derived cells examined. Contrastingly, the heterochromatin marker H3K9me3 (trimethylation of H3 lysine 9) mapped to all four HORs and ChIP–PCR showed an altered pattern of H3K9me3 in cancer cell lines and breast tumors, consistent with a reduction on the kinetochore-forming HORs. The JMJD2B demethylase is overexpressed in breast tumors with a CIN phenotype, and overexpression of exogenous JMJD2B in cultured breast epithelial cells caused loss of centromere-associated H3K9me3 and increased CIN. These findings suggest that impaired maintenance of pericentromeric heterochromatin may contribute to CIN in cancer and be a novel therapeutic target.

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Acknowledgements

We thank Kinya Yoda for advice with ChIP assays; Simone Schuffenhauer and Jan Murken for BF cells; Hunt Willard, Mariano Rocci and Theo Hulsebos for centromere plasmid probes; Tim Yen for CENP-E antibodies; Mervin Yoder for HUVEC samples; Virginia Thurston for advice on the patient BF sample and Andrew Ross, Nikki Collins, Janak Bhavsar, Tony Brown, Waylan Bessler and Laura Mead for technical assistance. This research was supported in part by INGEN (Indiana Genomics Initiative), the IUSM Cytogenetics Division and grants from the US DOD BCRP (W81XWH-09-1-0424) and American Cancer Society (IRG-84-002-24) awarded to BRG, IUSM Biomedical Research Grants awarded to RBS and an NCI grant (N01 CN-43300) to BSH. DS was supported by Deutsche Forschungsgemeinschaft and the Friedrich Baur Institute. INGEN is supported in part by Lilly Endowment. MR was supported by an NIH pre-doctoral fellowship (NRSA 1 T32 CA 111198 Cancer Biology Training Program).

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Author notes

  1. R J Hickey

    Present address: 5Current address: City of Hope, Duarte, CA, USA.,

  2. T Schwarz

    Present address: 6Current address: University of Wurzburg, Germany.,

  3. S Christan

    Present address: 7Current address: Technical University of Munich, Germany.,

  4. D Schindelhauer

    Present address: 8Current address: Chromosome Medicine Procurement, Therese-Studer-Str. 47, 80797 Munich, Germany.,

Authors and Affiliations

  1. Department of Medical and Molecular Genetics, Indiana University School of Medicine (IUSM), Indianapolis, IN, USA

    R B Slee, C M Steiner, B-S Herbert, G H Vance, M Radovich, B P Schneider & B R Grimes

  2. Indiana University Melvin and Bren Simon Cancer Center, Indianapolis, IN, USA

    R B Slee, B-S Herbert, G H Vance, R J Hickey, B P Schneider & B R Grimes

  3. Department of Medicine, IUSM, Indianapolis, IN, USA

    R J Hickey & B P Schneider

  4. Department of Medical Genetics, Ludwig Maximilians University, Munich, Germany

    T Schwarz, S Christan & D Schindelhauer

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  1. R B Slee
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Slee, R., Steiner, C., Herbert, BS. et al. Cancer-associated alteration of pericentromeric heterochromatin may contribute to chromosome instability. Oncogene 31, 3244–3253 (2012). https://doi.org/10.1038/onc.2011.502

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