Nucleic Acids Research, Vol 27, Issue 15 3229-3235, Copyright © 1999 by Oxford University Press
RO Pieper, KA Lester and CP Fanton
Growth constraint of bacterial and human cells has been shown to trigger
genetic mutation. We questioned whether growth constraint might also
trigger epigenetic mutation in the form of CpG island methylation.
Logarithmically growing normal human fibro-blasts (NHF) displayed little
(0-15%) CpG methylation in select regions of three CpG islands [estrogen
receptor (ER), E-cadherin (ECAD) and O (6)-methylguanine-DNA
methyltransferase (MGMT)] examined. NHF grown to and left at confluence for
2-21 days showed little (<10%) CpG methylation in the ER and ECAD CpG
islands. These confluent, growth-arrested cells, however, displayed
extensive ( approximately 50%) methylation of the MGMT CpG island. CpG
methylation in the MGMT CpG island was not associated with cellular
senescence. The methylation was, however, heritable, but not permanent, as
the level of CpG methylation in the MGMT CpG island of cells 4 population
doublings following replating after confluence were no different from those
in confluent cultures, but returned to levels noted in logarithmically
growing cells by 10 population doublings following replating. These results
suggest that growth constraint can trigger transient epigenetic change even
in normal non-senescent human cells.
ARTICLES
Confluence-induced alterations in CpG island methylation in cultured normal human fibroblasts
Department of Neurological Surgery and the UCSF Cancer Center, Room N261, 2340 Sutter Street, University of California-San Francisco, San Francisco, CA 94115-0128, USA. rpieper@cc.ucsf.edu
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