Bioluminescent imaging of TRAIL-induced apoptosis through detection of caspase activation following cleavage of DEVD-aminoluciferin.
Apoptosis, the most common and well-defined form of programmed cell death (PCD), is often impaired in cancer and neurodegenerative diseases and can limit conventional therapy. Bioluminescent molecular imaging was employed to study apoptosis in human colon cancer cells that have been treated with various doses of the therapeutic agent TRAIL (tumor necrosis factor-related apoptosis inducing ligand). While monitoring therapeutic response through a proluminescent, caspase-activated DEVD-aminoluciferin reagent (Caspase-Glo 3/7) which produced strong, stable signals, alternate preparations of the reagent were explored for non-invasive imaging methods. Dissolving the lyophilized DEVD-aminoluciferin compound in Dulbecco's PBS instead of lysis buffer (along with heat inactivation of an accompanying exogenous luciferase protein by heating at 85 degrees C for 20 minutes) yielded a minimally invasive apoptosis detector, with maximum luminescence intensities 2.5-fold stronger than those produced by D-luciferin at a final concentration of 100 microg/mL. Bioluminescent imaging of cancer therapeutic response through minimally invasive detection of caspase activation may serve as an important tool in monitoring apoptosis in vivo and in vitro.
Liu
Jue Judy
JJ
Roy and Diana Vagelos Scholars Program in Molecular Life Sciences, University of Pennsylvania School of Arts and Sciences, Philadelphia, USA.
Wang
Wenge
W
Dicker
David T
DT
El-Deiry
Wafik S
WS
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D051017:Apoptosis Regulatory Proteins / Q000494:pharmacology*; D001681:Biological Assay; D020169:Caspases / Q000378:metabolism*; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000175:diagnosis* / Q000201:enzymology / Q000473:pathology; D004789:Enzyme Activation; D049454:Firefly Luciferin / Q000031:analogs & derivatives* / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D049408:Luminescent Agents; D008163:Luminescent Measurements; D008562:Membrane Glycoproteins / Q000494:pharmacology*; D008853:Microscopy; D009842:Oligopeptides / Q000378:metabolism*; D053221:TNF-Related Apoptosis-Inducing Ligand; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D000970:Antineoplastic Agents; D051017:Apoptosis Regulatory Proteins; D049408:Luminescent Agents; D008562:Membrane Glycoproteins; D009842:Oligopeptides; D053221:TNF-Related Apoptosis-Inducing Ligand; C505653:TNFSF10 protein, human; D014409:Tumor Necrosis Factor-alpha; C511444:aspartyl-glutamyl-valyl-aspartyl-aminoluciferin; D049454:Firefly Luciferin; D020169:Caspases