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KMID : 0356720040200040191
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Journal of the Korean Society of Coloproctology
2004 Volume.20 No. 4 p.191 ~ p.198
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Sulindac-induced Apoptosis without Oligonucleosomal DNA Fragmentation in HT-29 Cells-Its Special References to Mitochondrial Pathway -
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Park Ki-Jae
Kwon Yuk
Kim Sung-Heun
Kim Min-Chan
Choi Hong-Jo
Kim Young-Hoon
Joh Se-Heon
Jung Ghap-Joong
Kim Sung-Hyun
Kwon Hyuk-Chan
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Abstract
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Purpose: This study was undertaken to reveal the molecular mechanism underlying sulindac-induced apoptosis in the human colon cancer cell line HT-29 (mutant p53).
Methods: Apoptosis was determined by using Hoechst 33342 staining, and translocation of proteins was established by using immunofluorescence, immunoelectron microscopy, and Western blotting after ultra- fractionation.
Results: This type of apoptosis was associated with decreased mitochondrial membrane potential, a translocation of the apoptosis-inducing factor (AIF) to the nucleus, and morphological evidence of nuclear condensation. However, DNA electrophoresis did not elucidate the ladder pattern of DNA fragments. Instead, a pulse-field gel electrophoresis showed that sulindac led to disintegration of nuclear DNA into-high- molecular-weight DNA fragments of about 100～300 kbp.
Conclusions: Our findings indicate that sulindac induces large-scale DNA fragmentation, suggesting a predominantly AIF-mediated cell-death process, through translocation of the AIF to the nucleus in HT-29 cells. J Korean Soc Coloproctol 2004;20:191-198
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KEYWORD
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Apoptosis, Colon cancer cell line HT-29, AIF, HMW DNA fragmentation
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