-
Original Article
-
J Korean Soc Coloproctol. 2004;20(4):191-198.
- Sulindac-induced Apoptosis without Oligonucleosomal DNA Fragmentation in HT-29 Cells: Its Special References to Mitochondrial Pathway.
-
Park, Ki Jae , Kwon, Yuk , Kim, Sung Heun , Kim, Min Chan , Choi, Hong Jo , Kim, Young Hoon , Cho, Se Heon , Jung, Ghap Joong , Kim, Sung Hyun , Kwon, Hyuk Chan
-
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.
Keywords :Apoptosis;Colon cancer cell line HT-29;AIF;HMW DNA fragmentation
