Changes in the protein spectrum of mitochondria isolated from hydroxycamptothecin-treated hepatoma cells.
As one of the most potent topoisomerase inhibitors, hydroxycamptothecin is more active and less toxic than conventional camptothecin. Recently, we found that hydroxycamptothecin can induce cell apoptosis via the mitochondrial pathway. This study was designed to investigate the mitochondrial protein profile in HCPT-treated cells using high-accuracy and high-sensitivity protein-identification technology. Of the 39 mitochondrial protein spots investigated, 25 displayed elevated and 14 suppressed abundance in hydroxycamptothecin-treated cells. The 25 spots were identified by mass spectrometry and they included proteins involved in many essential cellular functions. The potential role of these proteins in hydroxycamptothecin-mediated apoptosis is also discussed. This study has produced a short list of mitochondrial proteins that might hold the key to the mechanism by which hydroxycamptothecin induces mitochondrial dysfunction and cell apoptosis. It has laid the foundation for further elucidating the role of hydroxycamptothecin during apoptosis. Successful applications of multiple techniques including two-dimensional gel electrophoresis, matrix-assisted laser desorption ionization time-of-flight mass spectrometry and Western blot analysis have demonstrated that proteomic analyses provide appropriate approaches for understanding of the roles of anticancer drugs.
Fu
Yu-Rong
YR
Department of Laboratory Medicine, Key Laboratory of Laboratory Medical Diagnostics of Ministry of Education, Chongqing University of Medical Science, Chongqing, PRC.
Yi
Zheng-Jun
ZJ
Yan
Yu-Rong
YR
Qiu
Zong-Yin
ZY
D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D002166:Camptothecin / Q000031:analogs & derivatives* / Q000494:pharmacology; D045744:Cell Line, Tumor; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects*; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D024101:Mitochondrial Proteins / Q000096:biosynthesis*
D000972:Antineoplastic Agents, Phytogenic; D024101:Mitochondrial Proteins; C028098:10-hydroxycamptothecin; D002166:Camptothecin