TY - JOUR
T1 - Cisplatin-induced apoptosis in Hep3B cells
T2 - Mitochondria-dependent and -independent pathways
AU - Kim, Ji Su
AU - Lee, Jae Myun
AU - Chwae, Yong Joon
AU - Kim, Yeon Hyang
AU - Lee, Jung Hwan
AU - Kim, Kunhong
AU - Lee, Tae Ho
AU - Kim, Se Jong
AU - Park, Jeon Han
N1 - Funding Information:
We would like to thank Dr. Jean Y.J. Wang for p73-α and -β constructs, Dr. William G. Kaelin Jr. for p73 DN construct, and Dr. Xiaodong Wang for anti-Smac antibody. This research was supported by a grant from the Korea Science and Engineering Foundation (2000-2-20900-011-3).
PY - 2004/4/15
Y1 - 2004/4/15
N2 - Human hepatoma cell lines undergo apoptosis after treatment with cisplatin (CP), by mechanisms that are not fully understood, although our previous study demonstrated that Fas-dependent or -independent pathways are involved. To elucidate the mechanisms of CP-induced apoptosis in Hep3B cells, which are Fas- and p53-negative, we investigated mitochondria associated pathways, the involvement of NF-κB, and p73 activation. Results of Western blot and flow cytometry assay revealed that the translocation of Bax, resulted in the loss of mitochondrial membrane potential (Δφm) and the efflux of cytochrome c and of second mitochondria-derived activator of caspase/DIABLO from mitochondria into the cytosol. Caspase-3, -8 and -9 were activated by CP treatment, however, CP-induced apoptosis was not completely blocked by pretreating with the pan-caspase inhibitor, benzyloxycarbonyl- valinyl-alaninyl-aspartyl-(O-methyl)-fluoromethylketone, indicating that caspase-independent apoptotic pathways might also be involved. RNase protection assay confirmed that NF-κB downregulation leading to the suppression of its target genes, such as XIAP and TRAF2, and p73 accumulation were also observed in Hep3B cells treated with CP. CP-induced apoptosis was inhibited to some extent by transiently overexpressed p73 dominant negative and XIAP, but not by p73DN or XIAP alone. In conclusion, this study demonstrates that CP-induced apoptosis in Hep3B cells is associated with mitochondrial dysregulation, NF-κB downregulation and p73 accumulation.
AB - Human hepatoma cell lines undergo apoptosis after treatment with cisplatin (CP), by mechanisms that are not fully understood, although our previous study demonstrated that Fas-dependent or -independent pathways are involved. To elucidate the mechanisms of CP-induced apoptosis in Hep3B cells, which are Fas- and p53-negative, we investigated mitochondria associated pathways, the involvement of NF-κB, and p73 activation. Results of Western blot and flow cytometry assay revealed that the translocation of Bax, resulted in the loss of mitochondrial membrane potential (Δφm) and the efflux of cytochrome c and of second mitochondria-derived activator of caspase/DIABLO from mitochondria into the cytosol. Caspase-3, -8 and -9 were activated by CP treatment, however, CP-induced apoptosis was not completely blocked by pretreating with the pan-caspase inhibitor, benzyloxycarbonyl- valinyl-alaninyl-aspartyl-(O-methyl)-fluoromethylketone, indicating that caspase-independent apoptotic pathways might also be involved. RNase protection assay confirmed that NF-κB downregulation leading to the suppression of its target genes, such as XIAP and TRAF2, and p73 accumulation were also observed in Hep3B cells treated with CP. CP-induced apoptosis was inhibited to some extent by transiently overexpressed p73 dominant negative and XIAP, but not by p73DN or XIAP alone. In conclusion, this study demonstrates that CP-induced apoptosis in Hep3B cells is associated with mitochondrial dysregulation, NF-κB downregulation and p73 accumulation.
KW - CP
KW - Cisplatin
KW - Mitochondrial membrane potential
KW - PI
KW - PT
KW - Permeability transition
KW - Propidium iodide
KW - Second mitochondria-derived activator of caspase
KW - Smac
KW - z-VAD-fmk
KW - Δφ
UR - http://www.scopus.com/inward/record.url?scp=1642326520&partnerID=8YFLogxK
U2 - 10.1016/j.bcp.2003.12.013
DO - 10.1016/j.bcp.2003.12.013
M3 - Article
C2 - 15041463
AN - SCOPUS:1642326520
VL - 67
SP - 1459
EP - 1468
JO - Biochemical Pharmacology
JF - Biochemical Pharmacology
SN - 0006-2952
IS - 8
ER -