Abstract:
:Cholesterol and related compounds can give rise to oxygenated sterol molecules (oxysterols) which are potent regulators of lymphoid cell growth. Oxysterols added exogenously cause cell death of several lines of cultured cells, and on the basis of limited criteria, it has been suggested that this death is apoptosis. In the present study, we show definitive evidence that 25-hydroxycholesterol (25OHC) kills cells of the clone CEM-C7 by apoptosis and establish the temporal sequence of related cellular and biochemical events. Cell shrinkage was evident as early as 12 h, while cell death was not evident until after 24 h. It mounted rapidly after that, and by 72 h, virtually all cells were dead. Electron microscopic analysis shows that by 24 h after treatment and before the onset of cell death, early ultrastructural features typical of apoptosis were present. DNA breaks were detected by TUNEL assay prior to the onset of cell death. Two types of specific DNA pieces often associated with apoptosis were found as increasing numbers of cells died. DNA fragments of 300 and 50 kbp were not appreciable until 42 h, and internucleosomal cleavage was observed by 48 h after oxysterol addition. None of these effects were seen in an oxysterol-resistant CEM subclone, establishing the specificity for apoptosis of the biochemical and morphological events. z-VAD.FMK, a peptide inhibitor of ICE-related proteases delayed but did not prevent the apoptosis of CEM-C7 cells induced by 25OHC. The addition of mevalonate partially protected CEM-C7 cells from apoptosis but did not restore cell growth.
journal_name
Exp Cell Resjournal_title
Experimental cell researchauthors
Ayala-Torres S,Moller PC,Johnson BH,Thompson EBdoi
10.1006/excr.1997.3630subject
Has Abstractpub_date
1997-08-25 00:00:00pages
35-47issue
1eissn
0014-4827issn
1090-2422pii
S0014-4827(97)93630-2journal_volume
235pub_type
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journal_title:Experimental cell research
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