Sulindac activates nuclear translocation of AIF, DFF40 and endonuclease G but not induces oligonucleosomal DNA fragmentation in HT-29 cells.
Sulindac is one of the most widely studied nonsteroidal anti-inflammatory drugs in the prevention of colon cancer. Thus, from the viewpoint of colon cancer chemotherapy it is important to reveal the mechanism of sulindac-induced cell death. This study was undertaken to dissect the molecular mechanism underlying sulindac-induced apoptosis in human colon cancer cell line HT-29 (mutant p53), focusing on nuclear translocation of AIF, DFF and endonuclease G. On induction of apoptosis by sulindac, it was associated with decreased mitochondrial membrane potential, nuclear expression of active caspase-3, cleavage of poly(ADP-ribose) polymerase, translocation of mitochondrial proteins to the nucleus, and morphological evidence of nuclear condensation. However, sulindac led to only disintegration of nuclear DNA into high molecular weight DNA fragments of about 100-300 kbp as determined by a pulse-field gel electrophoresis, suggesting a predominantly AIF-mediated cell death process. In summary, our findings indicate that sulindac induces large-scale DNA fragmentation without oligonucleosomal DNA fragmentation. This result suggests that nuclear translocation of DFF and endonuclease G are not sufficient for the induction of oligonucleosomal DNA fragmentation in HT-29 cells.
Park
Young Chul
YC
Department of Anatomy and Cell Biology, Dong-A University College of Medicine (BK21 Program), Busan 602-714, South Korea.
Jeong
Jin Hee
JH
Park
Ki Jae
KJ
Choi
Hong Jo
HJ
Park
Yeong Min
YM
Jeong
Byung Kap
BK
Higuchi
Yoshihiro
Y
Yoo
Young Hyun
YH
D021581:Active Transport, Cell Nucleus / Q000187:drug effects*; D017209:Apoptosis / Q000187:drug effects*; D051033:Apoptosis Inducing Factor; D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D053938:DNA Fragmentation / Q000187:drug effects*; D003851:Deoxyribonucleases / Q000378:metabolism*; D016521:Electrophoresis, Gel, Pulsed-Field; D004706:Endodeoxyribonucleases / Q000378:metabolism*; D005420:Flavoproteins / Q000378:metabolism*; D019073:HT29 Cells; D006801:Humans; D020287:In Situ Nick-End Labeling; D008564:Membrane Potentials / Q000187:drug effects; D008565:Membrane Proteins / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D008928:Mitochondria / Q000502:physiology; D000075223:Poly-ADP-Ribose Binding Proteins; D013467:Sulindac / Q000494:pharmacology*
C494829:AIFM1 protein, human; D051033:Apoptosis Inducing Factor; D005420:Flavoproteins; D008565:Membrane Proteins; D000075223:Poly-ADP-Ribose Binding Proteins; D013467:Sulindac; C113696:DFFB protein, human; D003851:Deoxyribonucleases; D004706:Endodeoxyribonucleases; C052391:endonuclease G; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases