Non-invasive fluorescence imaging of cell death in fresh human colon epithelia treated with 5-Fluorouracil, CPT-11 and/or TRAIL.
Apoptosis is instrumental in several physiological/pathophysiological processes and is a frequently used end-point in the development of anti-neoplastic compounds. Despite ample data on several colon cancer cell lines, little is known about the susceptibility of human colon to apoptosis following treatment with established chemotherapeutics. By treating fresh human colonic explants with 5-Fluorouracil (200 microg/ml), CPT-11 (100 microg/ml) and/or TRAIL (100 ng/ml) we readily detected a signal in situ using FITC-VAD-FMK at different time points, whereas labeling of colonic explants with EGFP-conjugated Annexin V proved less specific. Although TRAIL treatment alone appeared to cause little apoptosis in human colonic epithelia versus the control, we observed a greater number of cells undergoing apoptosis when a combination of CPT-11 and TRAIL was used as compared to either agent alone. This is the initial demonstration of TRAIL-induced apoptosis with or without a chemotherapeutic agent in fresh primary human colon epithelia explants. Thus, human colonic explants may provide a valuable reference point when candidate therapeutic compounds triggering apoptosis in colon cancer cell lines, xenografts or mouse models are developed. The results support the feasibility of developing non-invasive optical imaging strategies to detect apoptosis through direct visualization of injury to human colonic epithelia in vivo.
Finnberg
Niklas
N
Abramson Comprehensive Cancer Center, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
Kim
Seok-Hyun
SH
Furth
Emma E
EE
Liu
Jue Judy
JJ
Russo
Pierre
P
Piccoli
David A
DA
Grimberg
Adda
A
El-Deiry
Wafik S
WS
D000293:Adolescent; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000494:pharmacology*; D002166:Camptothecin / Q000031:analogs & derivatives* / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003106:Colon / Q000166:cytology / Q000187:drug effects* / Q000601:surgery; D004338:Drug Combinations; D005260:Female; D005453:Fluorescence; D005472:Fluorouracil / Q000494:pharmacology*; D006801:Humans; D007091:Image Processing, Computer-Assisted; D000077146:Irinotecan; D008562:Membrane Glycoproteins / Q000494:pharmacology*; D009924:Organ Culture Techniques; D053221:TNF-Related Apoptosis-Inducing Ligand; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D051017:Apoptosis Regulatory Proteins; D004338:Drug Combinations; D008562:Membrane Glycoproteins; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; C505654:Tnfsf10 protein, mouse; D014409:Tumor Necrosis Factor-alpha; D000077146:Irinotecan; D005472:Fluorouracil; D002166:Camptothecin