Evaluation of the protective effects of all-trans-astaxanthin on canine osteosarcoma cell lines

Joseph J. Wakshlag Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Cheryl A. Balkman Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Stewart K. Morgan Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Margaret C. McEntee Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

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Abstract

Objective—To determine the effects of the antioxidant astaxanthin on growth of canine osteosarcoma cells with and without concurrent chemotherapeutic or irradiation insult.

Sample Population—Cells from 3 established canine osteosarcoma cell lines (D17, OS 2.4, and HMPOS).

Procedures—Growth-curve kinetics and cell cytotoxic effects were assessed by means of various treatment combinations and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Western blotting was performed to examine previously identified signaling pathways that astaxanthin reportedly affects. Additionally, cell-cycle kinetic evaluations, soft agar colony-forming assays, and antioxidant assays were performed to better understand the effect of astaxanthin on cell growth and function.

Results—Exposure to astaxanthin alone resulted in a mild to pronounced attenuation of cell proliferation in vitro, depending on the cell line, and did not interfere with the cell-death response to doxorubicin, irradiation, or peroxide-mediated insult. In some instances, astaxanthin acted in an additive fashion to augment cell death. Astaxanthin exposure increased the antioxidant potential of cells, whereas peroxide-mediated cell stress increased the antioxidant potential to the same degree as astaxanthin exposure or greater. No dramatic changes in phosphorylation of protein kinase B or upregulation of connexin 43 were detected.

Conclusions and Clinical Relevance—Findings suggested that astaxanthin administration may be beneficial in treatment of dogs for osteosarcoma. Its actions as an antioxidant did not improve osteosarcoma cell survival during chemotherapeutic or irradiation insults, warranting further research into this natural compound as an adjuvant, antiproliferative treatment for osteosarcoma in dogs.

Abstract

Objective—To determine the effects of the antioxidant astaxanthin on growth of canine osteosarcoma cells with and without concurrent chemotherapeutic or irradiation insult.

Sample Population—Cells from 3 established canine osteosarcoma cell lines (D17, OS 2.4, and HMPOS).

Procedures—Growth-curve kinetics and cell cytotoxic effects were assessed by means of various treatment combinations and a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Western blotting was performed to examine previously identified signaling pathways that astaxanthin reportedly affects. Additionally, cell-cycle kinetic evaluations, soft agar colony-forming assays, and antioxidant assays were performed to better understand the effect of astaxanthin on cell growth and function.

Results—Exposure to astaxanthin alone resulted in a mild to pronounced attenuation of cell proliferation in vitro, depending on the cell line, and did not interfere with the cell-death response to doxorubicin, irradiation, or peroxide-mediated insult. In some instances, astaxanthin acted in an additive fashion to augment cell death. Astaxanthin exposure increased the antioxidant potential of cells, whereas peroxide-mediated cell stress increased the antioxidant potential to the same degree as astaxanthin exposure or greater. No dramatic changes in phosphorylation of protein kinase B or upregulation of connexin 43 were detected.

Conclusions and Clinical Relevance—Findings suggested that astaxanthin administration may be beneficial in treatment of dogs for osteosarcoma. Its actions as an antioxidant did not improve osteosarcoma cell survival during chemotherapeutic or irradiation insults, warranting further research into this natural compound as an adjuvant, antiproliferative treatment for osteosarcoma in dogs.

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