YC-1 induces apoptosis of human renal carcinoma A498 cells in vitro and in vivo through activation of the JNK pathway
Journal
British Journal of Pharmacology
Journal Volume
155
Journal Issue
4
Pages
505-513
Date Issued
2008
Author(s)
Abstract
Background and purpose: The aim of this study was to elucidate the mechanism of YC-1{3-(5′-hydroxy methyl-2′-furyl)-1-benzylindazole}- induced human renal carcinoma cells apoptosis and to evaluate the potency of YC-1 in models of tumour growth in mice. Experimental approach: YC-1-mediated apoptosis was assessed by analysis of MTT, SRB, DAPI staining and flow cytometry analysis. Knockdown of JNK protein was achieved by transient transfection using siRNA. The mechanisms of action of YC-1 on different signalling pathways involved were studied using western blot. Fas clustering was analysed by confocal microscopy and in vivo efficacy was examined in a A498 xenograft model. Key results: YC-1 displayed cytotoxicity in renal carcinoma cells at 10 -7-10 -8 M. Increased condensation of chromatin was observed and an increase in the cell population in subG1 phase. Moreover, YC-1 triggered mitochondria-mediated and caspase-dependent pathways. YC-1 significantly induced Fas ligand expression, but did not modify either the protein levels of death receptors or ligands. In addition, Fas clustering in cells responsive to YC-1 was observed, suggesting involvement of a Fas-mediated pathway. Furthermore, YC-1 markedly induced phosphorylation of JNK and a JNK inhibitor, SP600125, and siRNA JNK1/2 significantly reversed YC-1-induced cytotoxicity and protein expression. We suggest that YC-1 induced JNK phosphorylation, the upregulation of FasL and Fas receptor clustering to promote the activation of caspases 8 and 3, resulting in apoptosis. Finally, we demonstrated the antitumour effect of YC-1 in vivo. Conclusions and implications: These data suggest that YC-1 is a good candidate for development as an anticancer drug. ? 2008 Macmillan Publishers Limited All rights reserved.
Subjects
Apoptosis; JNK; Renal cancer; YC-1
SDGs
Other Subjects
1 benzyl 3 (5 hydroxymethyl 2 furyl)indazole; anthra[1,9 cd]pyrazol 6(2h) one; caspase 3; caspase 8; Fas antigen; Fas ligand; small interfering RNA; stress activated protein kinase inhibitor; animal experiment; animal model; animal tissue; antineoplastic activity; apoptosis; article; cancer cell culture; confocal microscopy; controlled study; drug dose comparison; human; human cell; kidney carcinoma; male; mouse; nonhuman; nucleotide sequence; priority journal; tumor xenograft; Animals; Antigens, CD95; Apoptosis; Carcinoma, Renal Cell; Caspase 3; Caspase 8; Chromatin; Dose-Response Relationship, Drug; Enzyme Activators; Fas Ligand Protein; G1 Phase; Humans; Indazoles; JNK Mitogen-Activated Protein Kinases; Male; Mice; Mice, Inbred BALB C; Mice, Nude; Phosphorylation
Type
journal article