J Korean Soc Coloproctol.  2004 Aug;20(4):191-198.

Sulindac-induced Apoptosis without Oligonucleosomal DNA Fragmentation in HT-29 Cells: Its Special References to Mitochondrial Pathway

Affiliations
  • 1Department of Surgery, Dong-A University College of Medicine, Busan, Korea. colonch@donga.ac.kr
  • 2Department of Internal Medicine, Dong-A University College of Medicine, Busan, Korea.

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.

Keyword

Apoptosis; Colon cancer cell line HT-29; AIF; HMW DNA fragmentation

MeSH Terms

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
Apoptosis Inducing Factor
DNA
Sulindac
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