Cisplatin-induced apoptosis in Hep3B cells

Mitochondria-dependent and -independent pathways

Ji Su Kim, Jae Myun Lee, Yong Joon Chwae, Yeon Hyang Kim, Jung Hwan Lee, Kunhong Kim, Tae Ho Lee, Se Jong Kim, Jeon Han Park

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1459-1468
Number of pages10
JournalBiochemical Pharmacology
Volume67
Issue number8
DOIs
Publication statusPublished - 2004 Apr 15

Fingerprint

Mitochondria
Cisplatin
Apoptosis
Caspases
Assays
Down-Regulation
TNF Receptor-Associated Factor 2
Caspase Inhibitors
Flow cytometry
Caspase 8
Mitochondrial Membrane Potential
Ribonucleases
Cytochromes c
Caspase 3
Cytosol
Hepatocellular Carcinoma
Flow Cytometry
Genes
Western Blotting
Chemical activation

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Pharmacology

Cite this

Kim, J. S., Lee, J. M., Chwae, Y. J., Kim, Y. H., Lee, J. H., Kim, K., ... Park, J. H. (2004). Cisplatin-induced apoptosis in Hep3B cells: Mitochondria-dependent and -independent pathways. Biochemical Pharmacology, 67(8), 1459-1468. https://doi.org/10.1016/j.bcp.2003.12.013
Kim, Ji Su ; Lee, Jae Myun ; Chwae, Yong Joon ; Kim, Yeon Hyang ; Lee, Jung Hwan ; Kim, Kunhong ; Lee, Tae Ho ; Kim, Se Jong ; Park, Jeon Han. / Cisplatin-induced apoptosis in Hep3B cells : Mitochondria-dependent and -independent pathways. In: Biochemical Pharmacology. 2004 ; Vol. 67, No. 8. pp. 1459-1468.
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abstract = "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.",
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Kim, JS, Lee, JM, Chwae, YJ, Kim, YH, Lee, JH, Kim, K, Lee, TH, Kim, SJ & Park, JH 2004, 'Cisplatin-induced apoptosis in Hep3B cells: Mitochondria-dependent and -independent pathways', Biochemical Pharmacology, vol. 67, no. 8, pp. 1459-1468. https://doi.org/10.1016/j.bcp.2003.12.013

Cisplatin-induced apoptosis in Hep3B cells : Mitochondria-dependent and -independent pathways. / Kim, Ji Su; Lee, Jae Myun; Chwae, Yong Joon; Kim, Yeon Hyang; Lee, Jung Hwan; Kim, Kunhong; Lee, Tae Ho; Kim, Se Jong; Park, Jeon Han.

In: Biochemical Pharmacology, Vol. 67, No. 8, 15.04.2004, p. 1459-1468.

Research output: Contribution to journalArticle

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

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.

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