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Aflatoxin-transformed C3H/10T1/2 Cells Overexpress Protein Kinase C and Have an Altered Response to Phorbol Ester Treatments

Joseph A. Dunn, Michael B. Faletto, Steven J. Kasper and Hira L. Gurtoo
Joseph A. Dunn
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Michael B. Faletto
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Steven J. Kasper
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Hira L. Gurtoo
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DOI:  Published February 1992
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Abstract

The aflatoxin B1-transformed C3H/10T1/2; (10T1/2) cell line 7SA has disordered growth in culture and is tumorigenic in syngeneic mice. Chronic exposure (14 days) of 10T1/2 and 7SA cells to phorbol 12,13-dibutyrate (PDBu) increased the saturation density of 10T1/2 cells but dramatically slowed the entry of 7SA cells into the log phase of growth without affecting their final saturation density. Similar PDBu treatment of low-density cultures dramatically decreased the size of 7SA colonies. Both cell lines bound [3H]PDBu in a specific and saturable manner. Analysis of this binding yielded linear Scatchard plots for both cell lines with distinctly different Kd values (10.7 nm for 10T1/2 versus 54.5 nm for 7SA). The total amount of [3H]PDBu bound was 2-fold greater in the 7SA cells versus the 10T1/2 cell line. Both cell lines released arachidonic acid following a 2-h exposure to PDBu; however, the magnitude of the response of the 7SA cells was only one-half that of the 10T1/2 cells. Western blot analysis of protein kinase C (PKC) using specific anti-PKC antibodies revealed a greater total amount of PKCα in the 7SA cells relative to an equal number of 10T1/2 cells. No immunoreactive PKCα was found in either cell line 16 h after exposure to 600 nm PDBu; however, PKCα returned to control levels in both cell lines 24 h after removal of the phorbol ester. These results suggest that an overexpression of PKCα may play a role in the altered biological properties of aflatoxin-transformed 10T1/2 cells.

Footnotes

  • ↵3 To whom all requests for reprints should be addressed, at present address: Burroughs Wellcome Company, 3030 Cornwallis Road, Research Triangle Park, NC 27709.

  • Received July 23, 1991.
  • Accepted November 21, 1991.
  • ©1992 American Association for Cancer Research.
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February 1992
Volume 52, Issue 4
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Aflatoxin-transformed C3H/10T1/2 Cells Overexpress Protein Kinase C and Have an Altered Response to Phorbol Ester Treatments
Joseph A. Dunn, Michael B. Faletto, Steven J. Kasper and Hira L. Gurtoo
Cancer Res February 15 1992 (52) (4) 990-996;

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Aflatoxin-transformed C3H/10T1/2 Cells Overexpress Protein Kinase C and Have an Altered Response to Phorbol Ester Treatments
Joseph A. Dunn, Michael B. Faletto, Steven J. Kasper and Hira L. Gurtoo
Cancer Res February 15 1992 (52) (4) 990-996;
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