Sulindac-induced Apoptosis without Oligonucleosomal DNA Fragmentation in HT-29 Cells: Its Special References to Mitochondrial Pathway.
- Author:
Ki Jae PARK
1
;
Yuk KWON
;
Sung Heun KIM
;
Min Chan KIM
;
Hong Jo CHOI
;
Young Hoon KIM
;
Se Heon CHO
;
Ghap Joong JUNG
;
Sung Hyun KIM
;
Hyuk Chan KWON
Author Information
1. Department of Surgery, Dong-A University College of Medicine, Busan, Korea. colonch@donga.ac.kr
- Publication Type:Original Article
- Keywords:
Apoptosis;
Colon cancer cell line HT-29;
AIF;
HMW DNA fragmentation
- MeSH:
Apoptosis Inducing Factor;
Apoptosis*;
Blotting, Western;
Cell Line;
Colonic Neoplasms;
DNA Fragmentation*;
DNA*;
Electrophoresis;
Fluorescent Antibody Technique;
HT29 Cells*;
Humans;
Membrane Potential, Mitochondrial;
Microscopy, Immunoelectron;
Sulindac
- From:Journal of the Korean Society of Coloproctology
2004;20(4):191-198
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
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.
