Simultaneous quantitative monitoring of drug-induced caspase cascade pathways in carcinoma cells.
Caspases are the key mediators of apoptosis. The caspase cascade includes a series of events leading to the activation of initiator and downstream caspases in a cell. Analysis of the caspase cascade in intact cells, however, has generally been limited as the simultaneous monitoring of upstream and downstream caspases is not well executed. In an effort to monitor the activation of caspase cascades in an intact cell, high-content cellular imaging that allows simultaneous quantitative monitoring of caspase activation has been developed. This has great significance for the exploration of various cellular caspases involved in apoptotic pathways as possible therapeutic targets in the process of drug discovery. To explore the potential of simultaneous monitoring of caspase-mediated apoptotic pathways, human myeloid leukemia HL-60 cells were treated with SH-03 {(7S,7aR,13aS)-9,10-dimethoxy-3,3-dimethyl-7,7a,13,13a-tetrahydro-3H-chromeno [3,4-b]pyrano[2,3-h]chromen-7-ol} (a newly synthesized candidate), camptothecin or naringenin (agents known to induce apoptosis) with or without caspase inhibitors. SH-03 or naringenin treatment initiated the caspase cascade through an intrinsic apoptotic pathway, whereas camptothecin treatment triggered both intrinsic and extrinsic caspase cascades. We now report a new approach based on uniform threshold intensity distribution that facilitates rapid, quantitative monitoring of drug-induced caspase cascades through multi-spectral and multicolor imaging cytometry.
Naoghare
Pravin K
PK
Research Institute of Pharmaceutical Sciences and College of Pharmacy, Seoul National University, Seoul 151-742, South Korea.
Ki
Hyeon A
HA
Paek
Seung-Mann
SM
Tak
Yu Kyung
YK
Suh
Young-Ger
YG
Kim
Sang Geon
SG
Lee
Kyeong-Hee
KH
Song
Joon Myong
JM
D000970:Antineoplastic Agents / Q000008:administration & dosage*; D002277:Carcinoma / Q000188:drug therapy* / Q000378:metabolism*; D061945:Caspase Inhibitors; D020169:Caspases / Q000378:metabolism*; D045744:Cell Line, Tumor; D003198:Computer Simulation; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D008954:Models, Biological; D015398:Signal Transduction / Q000187:drug effects*
D000970:Antineoplastic Agents; D061945:Caspase Inhibitors; D020169:Caspases