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  • Original Paper
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Serine protease inhibitor TPCK prevents Taxol-induced cell death and blocks c-Raf-1 and Bcl-2 phosphorylation in human breast carcinoma cells

Oncogene volume 18, pages 3431–3439 (1999)Cite this article

Abstract

The mechanism of Taxol-induced apoptosis was investigated in MCF-7 human breast carcinoma cells. Taxol-induced apoptosis was associated with phosphorylation of both c-Raf-1 and Bcl-2 and activation of ERK and JNK MAP kinases. The serine protease inhibitor N-tosyl-L-phenylalanine chloromethyl ketone (TPCK) effectively blocked apoptosis, but N-p-tosyl-L-lysine chloromethyl ketone (TLCK), another serine protease inhibitor, was without effect. TPCK treatment also prevented phosphorylation of c-Raf-1 and Bcl-2 in response to Taxol treatment. The serine protease inhibitor did not alter JNK activity, but it enhanced Taxol-induced activation of ERK1/2. Treatment of cells with the inhibitor of MEK activation, PD98059, prevented Taxol-induced ERK activation both in the presence and absence of TPCK, but did not influence survival of either Taxol- or Taxol plus TPCK-treated cells. In addition, PD98059 had no effect on c-Raf-1 or Bcl-2 phosphorylation. Thus, while the Taxol-induced phosphorylations of c-Raf-1 and Bcl-2 proteins appear to be coupled, these events can be disassociated from ERK1/2 activation. In summary, these findings suggest that phosphorylation of c-Raf-1 and Bcl-2, but not ERK1/2, are important signaling events in Taxol-induced apoptosis of MCF-7 breast cancer cells and that a TPCK inhibitable protease(s) is required for these processes.

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Abbreviations

ERK:

extracellular signal regulated kinase

JNK:

c-Jun N-terminal protein kinase

TPCK:

N-tosyl-L-phenylalanine chloromethyl ketone

TLCK:

N-p-tosyl-L-lysine chloromethyl ketone

DAPI:

4,6,-diamidino-2 phenyl-indole

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Authors and Affiliations

  1. Gene Expression and Aging Section, Laboratory of Biological Chemistry, National Institute on Aging, Baltimore, 21224, Maryland, MD, USA

    Ying Huang & Nikki J Holbrook

  2. Division of Basic Sciences, National Cancer Institute, Bethesda, 20892, Maryland, MD, USA

    M Saeed Sheikh & Albert J Fornace Jr

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  1. Ying Huang
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  2. M Saeed Sheikh
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Huang, Y., Sheikh, M., Fornace, A. et al. Serine protease inhibitor TPCK prevents Taxol-induced cell death and blocks c-Raf-1 and Bcl-2 phosphorylation in human breast carcinoma cells. Oncogene 18, 3431–3439 (1999). https://doi.org/10.1038/sj.onc.1202685

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