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Differentiation of pancreatic ductal carcinoma cells associated with selective expression of protein kinase C isoforms

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Abstract

Background: The signal transduction pathways important in regulating the growth and differentiation of malignant cells are poorly understood. Recent evidence has implicated activation of the protein kinase C (PKC) family of signaling proteins in pancreatic carcinoma during cytokine-induced cytostasis and differentiation.

Methods: A human pancreatic adenocarcinoma (HPAC) cell line was exposed to tumor necrosis factor-α (TNF-α; 40 ng/ml) for 6 days. Cytostasis and viability were confirmed by daily MTT [(3(4,5)-dimethyl-thiazol-2-yl) 2,5-diphenyl-tetrazolium bromide] and trypan exclusion assay. Protein fractions were isolated daily and subjected to immunoblot analysis for the normal (terminally differentiated) pancreatic ductal cell marker carbonic anhydrase II (CA II) as well as specific PKC isoforms (α, β, γ, η, andζ).

Results: Growth arrest occurred in HPAC cells after exposure to TNF-α for 48 h, with viability maintained above 90% throughout the 6-day time course. CA II immunoreactivity was not detected in untreated controls but appeared after 2 days of TNF-α exposure, peaking on day 6. Concurrently, TNF-α induced the selective downregulation of PKC-α, whereas PKC-γ levels increased. PKC-β and PKC-η immunoreactivity did not change. The atypical PKC-ζ isoform developed a doublet banding pattern in response to TNF-α, although overall PKC-ζ levels did not change.

Conclusions: TNF-α-induced growth arrest and differentiation in HPAC cells is associated with the selective downregulation of PKC-α and upregulation of PKC-γ.

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Authors and Affiliations

  1. From the Department of Surgery, University of South Florida College of Medicine, Tampa, Florida, U.S.A.

    Michael G. Franz MD, James G. Norman MD & Peter J. Fabri MD

  2. the Department of Biochemistry and Molecular Biology, University of South Florida College of Medicine, Tampa, Florida, U.S.A.

    William R. Gower Jr PhD

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  1. Michael G. Franz MD
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  2. James G. Norman MD
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  3. Peter J. Fabri MD
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  4. William R. Gower Jr PhD
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Franz, M.G., Norman, J.G., Fabri, P.J. et al. Differentiation of pancreatic ductal carcinoma cells associated with selective expression of protein kinase C isoforms. Annals of Surgical Oncology 3, 564–569 (1996). https://doi.org/10.1007/BF02306090

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  • DOI: https://doi.org/10.1007/BF02306090

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