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Sensitization of human breast cancer cells to gemcitabine by the protein kinase C modulator bryostatin 1

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Abstract

Purpose

Protein kinase C (PKC) plays an important role in cell proliferation, differentiation, and apoptosis. The interaction between the PKC modulator bryostatin 1 (BRYO), and gemcitabine in human breast cancer MCF-7 and MDA-MB-231 cells and in the non-transformed MCF-10A human breast epithelial cells was investigated.

Methods

Immunoblotting was used to determine the expression of PKC isoenzymes and proteins involved in the cell cycle and apoptosis. MTT, ELISA and flow cytometry assays were used to determine cell survival.

Results

Treatment of cells with BRYO 200 nM resulted in a significant downregulation of cytoplasmic PKC in all three cell lines. However, the expression of membranous PKC was differentially affected in these cells. BRYO (1–200 nM) had no significant effects on cell viability in any of the cell lines. Nevertheless, BRYO significantly enhanced the antiproliferative and apoptotic effects of gemcitabine in the MCF-7 and MDA-MB-231 cells, but not in the MCF-10A cells. This was associated with significant reduction in the bcl-2/bax ratio. There was a significant upregulation of p53, p21waf1, and p27 in MCF7 and MCF-10A cells treated with the combination of gemcitabine and BRYO compared to gemcitabine-treated cells.

Conclusions

The potentiation of the effect of gemcitabine by BRYO was demonstrated in MCF-7 and MDA-MB-231 cells and was associated with a specific pattern of PKC modulation. Further investigation of the role of specific isoforms of PKC in the downstream molecular events of gemcitabine-induced cytotoxicity is warranted.

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Authors and Affiliations

  1. Karmanos Cancer Institute, Wayne State University, Detroit, MI 48201, USA

    Shadan Ali, Olivia Aranha, Yiwei Li, Fazlul H. Sarkar & Philip Agop Philip

  2. The Institute of Cancer Research, Arizona State University, Tempe, AZ, USA

    George R. Pettit

  3. Division of Hematology and Oncology, Barbara Ann Karmanos Cancer Institute, Wayne State University, 501 Hudson Building, 3990 John R Street, Detroit, MI 48201, USA

    Philip Agop Philip

Authors
  1. Shadan Ali
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  2. Olivia Aranha
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  3. Yiwei Li
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  4. George R. Pettit
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  5. Fazlul H. Sarkar
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  6. Philip Agop Philip
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Correspondence to Philip Agop Philip.

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Ali, S., Aranha, O., Li, Y. et al. Sensitization of human breast cancer cells to gemcitabine by the protein kinase C modulator bryostatin 1. Cancer Chemother Pharmacol 52, 235–246 (2003). https://doi.org/10.1007/s00280-003-0628-6

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  • DOI: https://doi.org/10.1007/s00280-003-0628-6

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