Induction of colon cancer cell death by 7-hydroxystaurosporine (UCN-01) is associated with increased p38 MAPK and decreased Bcl-xL.
UCN-01, a selective inhibitor of protein kinase C, is known to inhibit the growth of cancer cells. Although it is currently undergoing clinical evaluation, information about its effect on human colon cancer is limited and the mechanism responsible is lacking. The objective of this study was to examine the cytotoxicity of UCN-01 to human colon cancer cells in vitro and its effect on the apoptotic molecules. HT-29, a radiation- and chemotherapy-resistant human colon cancer cell, was used in the study. Cell death/apoptosis was determined by the MTT assay and DNA fragmentation measurement. NF-kappaB activity was measured by an enzyme immunoassay method. Western blot was employed to examine the expression of relevant apoptotic molecules. The result showed that UCN-01 could induce apoptosis of human colon cancer cells in a time- and dose-dependent manner. It markedly reduced the expression of Bcl-xL, but enhanced the level of p38 MAPK. In addition to Bcl-xL and p38 MAPK, UCN-01 also increased both caspase-3 and peroxisome proliferator activated receptor gamma protein levels. HT-29 cells transfected with exogenous Bcl-xL showed a significant increase in NF-kappaB activity and prevented apoptosis induced by UCN-01. The overexpression of Bcl-xL also reversed other relevant molecular changes observed in UCN-01-treated cells. In conclusion, UCN-01 exerted an antitumor effect in human colon cancer cells by inducing apoptosis. The mechanism responsible appeared to be related to reduction of Bcl-xL and increased p38 MAPK. The overexpression of Bcl-xL can significantly prevent apoptosis induced by UCN-01.
Chan
Ursula P F
UP
Department of Surgery, Prince of Wales Hospital, The Chinese University of Hong Kong.
Lee
Janet F Y
JF
Wang
S H
SH
Leung
Ka L
KL
Chen
George G
GG
D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3; D020169:Caspases / Q000096:biosynthesis; D003110:Colonic Neoplasms; D004305:Dose-Response Relationship, Drug; D019073:HT29 Cells; D006801:Humans; D020928:Mitogen-Activated Protein Kinases / Q000096:biosynthesis; D016328:NF-kappa B / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000037:antagonists & inhibitors; D018160:Receptors, Cytoplasmic and Nuclear / Q000096:biosynthesis; D019311:Staurosporine / Q000031:analogs & derivatives* / Q000494:pharmacology*; D014157:Transcription Factors / Q000096:biosynthesis; D051020:bcl-X Protein; D048051:p38 Mitogen-Activated Protein Kinases
C494811:BCL2L1 protein, human; D016328:NF-kappa B; D019253:Proto-Oncogene Proteins c-bcl-2; D018160:Receptors, Cytoplasmic and Nuclear; D014157:Transcription Factors; D051020:bcl-X Protein; C054852:7-hydroxystaurosporine; D011493:Protein Kinase C; D020928:Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D019311:Staurosporine