5-Lipoxygenase expression and tepoxalin-induced cell death in squamous cell carcinomas in cats

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

Search for other papers by Joseph J. Wakshlag in
Current site
Google Scholar
PubMed
Close
 DVM, PhD
,
Jeanine Peters-Kennedy Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

Search for other papers by Jeanine Peters-Kennedy in
Current site
Google Scholar
PubMed
Close
,
Jennifer J. Bushey Departments of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

Search for other papers by Jennifer J. Bushey in
Current site
Google Scholar
PubMed
Close
 DVM
, and
John P. Loftus Departments of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY 14853.

Search for other papers by John P. Loftus in
Current site
Google Scholar
PubMed
Close
 PhD

Abstract

Objective—To assess expression pattern and subcellular compartmentalization of 5-lipoxygenase in cutaneous, UV radiation–induced, and oral squamous cell carcinomas (SCCs) in cats and determine the effects of cyclooxygenase or 5-lipoxygenase inhibition on proliferation or apoptosis in a feline oral squamous cell carcinoma (SCCF1) cell line.

Sample—60 archived paraffin-embedded samples of SCCs from 60 cats and SCCF1 cells.

Procedures—Retrospective immunohistochemical analysis of the archived samples of SCCs (20 cutaneous, 20 UV radiation–induced, and 20 oral tumors) was performed. Cell culture proliferation assays involving SCCF1 cells were performed, and tepoxalin-induced apoptosis and signaling were examined via western blotting and annexin V staining.

Results—Immunohistochemically, staining for 5-lipoxygenase was most frequently of greatest intensity in oral SCCs, whereas staining of cutaneous and UV radiation–induced lesions had less consistent 5-lipoxygenase expression. Exposure of SCCF1 cells to the 5-lipoxygenase inhibitor tepoxalin resulted in apoptosis; the effect appeared to be mediated via alteration of cell signaling rather than via suppression of lipid mediators that are typically produced as a result of 5-lipoxygenase activity.

Conclusions and Clinical Relevance—In cats, expression of 5-lipoxygenase in SCCs appeared to differ depending on tumor location. The influence of tepoxalin-induced 5-lipoxygenase inhibition on a 5-lipoxygenase–expressing cell line coupled with the notable expression of 5-lipoxygenase in oral SCCs suggested that 5-lipoxygenase inhibition may have therapeutic benefits in affected cats. Although the safety of tepoxalin in cats has yet to be investigated, 5-lipoxygenase inhibitors should be evaluated for use as a potential treatment for SCCs in that species.

Abstract

Objective—To assess expression pattern and subcellular compartmentalization of 5-lipoxygenase in cutaneous, UV radiation–induced, and oral squamous cell carcinomas (SCCs) in cats and determine the effects of cyclooxygenase or 5-lipoxygenase inhibition on proliferation or apoptosis in a feline oral squamous cell carcinoma (SCCF1) cell line.

Sample—60 archived paraffin-embedded samples of SCCs from 60 cats and SCCF1 cells.

Procedures—Retrospective immunohistochemical analysis of the archived samples of SCCs (20 cutaneous, 20 UV radiation–induced, and 20 oral tumors) was performed. Cell culture proliferation assays involving SCCF1 cells were performed, and tepoxalin-induced apoptosis and signaling were examined via western blotting and annexin V staining.

Results—Immunohistochemically, staining for 5-lipoxygenase was most frequently of greatest intensity in oral SCCs, whereas staining of cutaneous and UV radiation–induced lesions had less consistent 5-lipoxygenase expression. Exposure of SCCF1 cells to the 5-lipoxygenase inhibitor tepoxalin resulted in apoptosis; the effect appeared to be mediated via alteration of cell signaling rather than via suppression of lipid mediators that are typically produced as a result of 5-lipoxygenase activity.

Conclusions and Clinical Relevance—In cats, expression of 5-lipoxygenase in SCCs appeared to differ depending on tumor location. The influence of tepoxalin-induced 5-lipoxygenase inhibition on a 5-lipoxygenase–expressing cell line coupled with the notable expression of 5-lipoxygenase in oral SCCs suggested that 5-lipoxygenase inhibition may have therapeutic benefits in affected cats. Although the safety of tepoxalin in cats has yet to be investigated, 5-lipoxygenase inhibitors should be evaluated for use as a potential treatment for SCCs in that species.

Contributor Notes

Supported by the Winn Feline Foundation.

Address correspondence to Dr. Wakshlag (dr.joesh@gmail.com).
  • 1.

    Vail DM, Withrow SJ. Tumors of the skin and subcutaneous tissues. In: Withrow SJ, Vail DM, eds. Small animal clinical oncology. 4th ed. St Louis: Saunders, 2007;375401.

    • Search Google Scholar
    • Export Citation
  • 2.

    Head KW, Else RW, Dubielzieg RR. Tumors of the alimentary tract. In: Meuten DJ, ed. Tumors in domestic animals. 4th ed. Ames, Iowa: Blackwell Publishing, 2002;401482.

    • Search Google Scholar
    • Export Citation
  • 3.

    Gil Z, Fiss DM. Contemporary management of head and neck tumors. Isr Med Assoc J 2009; 11: 296300.

  • 4.

    Pannone G, Bufo P, Grothey A, et al. Cyclooxygenase-2 expression in oral squamous cell carcinoma. Int J Immunopath Pharmacol 2004; 17: 273282.

  • 5.

    Pestili de Almeida EM, Piche C, Sirios J, et al. Expression of cyclo-oxygenase 2 in naturally occurring squamous cell carcinomas in dogs. J Histochem Cytochem 2001; 49: 867875.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 6.

    Mohammed SI, Khan KN, Sellers RS, et al. Expression of cyclooxygenase-1 and 2 in naturally-occurring canine cancer. Prostaglandins Leukot Essent Fatty Acids 2004; 70: 479483.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 7.

    Hayes A, Scase J, Miller S, et al. COX-1 and COX-2 expression in feline oral squamous cell carcinoma. J Comp Pathol 2006; 135: 9399.

  • 8.

    DiBernardi L, Dore M, Davis JA, et al. Study of feline oral squamous cell carcinoma: potential target for cyclooxygenase inhibitor treatment. Prostaglandins Leukot Essent Fatty Acids 2007; 76: 245250.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 9.

    Heller DA, Fan TM, deLorimier LP et al. In vitro cyclooxygense-2 protein expression and enzymatic activity in neoplastic cells. J Vet Intern Med 2007; 21: 10481055.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 10.

    Li N, Sood S, Wang S, et al. Overexpression of 5-lipoxygenase and cyclooxygenase 2 in hamster and human oral cancer and chemopreventative effects of Zileuton and Celecoxib. Clin Cancer Res 2005; 11: 20892096.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 11.

    Sun Z, Sood S, Li N, et al. Involvement of the 5-lipoxygenase/leukotriene A4 hydrolase pathway in 7,12-DMBA-induced oral carcinogenesis in hamster cheek pouch and inhibition of carcinogenesis by its inhibitors. Carcinogenesis 2006; 27: 19021908.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 12.

    Yang P, Sun Z, Chan D, et al. Zyflamend reduces LTB4 formation and prevents oral carcinogenesis in a DMBA-induced hamster cheek pouch model. Carcinogenesis 2008; 29: 21822189.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 13.

    Schroeder CP, Yang P, Newman RA, et al. Eicosanoid metabolism in squamous cell carcinoma cell lines derived from primary and metastatic head and neck cancer and its modulation by Celecoxib. CurrBiol Ther 2004; 9: 847852.

    • Search Google Scholar
    • Export Citation
  • 14.

    Porteder H, Matejka M, Ulrish W, et al. The cyclo-oxygenase and lipoxygenase pathways in human oral cancer tissue. J Maxillofac Surg 1984; 12: 145147.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 15.

    Metzger K, Angres G, Maier H, et al. Lipoxygenase products in human saliva: patients with oral cancer compared to controls. Free Radic Biol Med 1995; 18: 185192.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 16.

    Duffield Lillico AJ, Boyle JO, Zhou XK, et al. Levels of prostaglandin E metabolite and leukotriene E4 are increased in the urine of smokers: evidence that celecoxib shunts arachidonic acid into the 5-lipoxygenase pathway. Cancer Prev Res 2009; 2: 322329.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 17.

    Furstenberger G, Krieg P, Muller-Decker K, et al. What are cyclooxygenases and lipoxygenases doing in the driver's seat of carcinogenesis. Int J Cancer 2006; 119: 22472254.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 18.

    O'Flaherty JT, Rogers LC, Pauma CM, et al. 5-oxo-ETE analogs and the proliferation of cancer cells. Biochim Biophys Acta 2005; 1736: 228236.

  • 19.

    Grant GE, Rokach J, Powell WS. 5-oxo-ETE and the OXE receptor. Prostaglandins Other Lipid Med 2009; 89: 98104.

  • 20.

    Rovati GE, Capra V. Cysteinyl-leukotriene receptors and cellular signals. Sci World J 2007; 7: 13751392.

  • 21.

    Lascelles BDX, Court MH, Hardle EM, et al. Nonsteroidal anti-inflammatory drugs in cats: a review. Vet Anesth Analg 2007; 34: 228250.

  • 22.

    Wakshlag JJ, McNeill CJ, Antonyak MA, et al. Expression and activity of transglutaminase II in spontaneous tumors of dogs and cats. J Comp Pathol 2006; 134: 202210.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 23.

    Tannehill-Gregg S, Kergosein E, Rosol TJ. Feline head and neck squamous cell carcinoma cell line: characterization, production of parathyroid hormone-related protein and regulation by transforming growth factor-β. In Vitro Cell Dev Biol 2001; 37: 676683.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 24.

    Wakshlag JJ, Antonyak MA, Boehm JE, et al. Effects of tissue transglutaminase on beta amyloid 1–42 induced apoptosis. Protein J 2006; 25: 8394.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 25.

    Antonyak MA, Miller AM, Jansen JM, et al. Augmentation of tissue transglutaminase expression and activation by epidermal growth factor inhibit doxorubicin-induced apoptosis in human breast cancer cells. J Biol Chem 2004; 279: 4146141467.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 26.

    Lin DT, Subbaramaiah K, Shah JP, et al. Cyclooxygenase-2: a novel molecular target for the prevention and treatment of head and neck cancer. Head Neck 2002; 24: 792799.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 27.

    Fecker LF, Stockfleth E, Nidl I, et al. The role of apoptosis in therapy and prophylaxis of epithelial tumours by nonsteroidal anti-inflammatory drugs (NSAIDs). BrJDermatol 2007; 156: 2533.

    • Search Google Scholar
    • Export Citation
  • 28.

    Ding XZ, Talamonti MS, Bell RH, et al. A novel anti-pancreatic cancer agent, LY293111. Anticancer Drugs 2005; 16: 467473.

  • 29.

    Radmark O, Werz O, Steinhiler D, et al. 5-lipoxygenase: regulation of expression and enzyme activity. Trends Biochem Sci 2007; 32: 332341.

  • 30.

    Covey TM, Edes K, Fitzpatrick FA. Akt activation by arachidonic acid metabolism occurs via oxidation and inactivation of PTEN tumor suppressor. Oncogene 2007; 26: 57845792.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 31.

    Giannoni E, Fiaschi T, Ramponi G, et al. Redox regulation of anoikis resistance of metastatic prostate cancer cells: key role for Src and EGFR-mediated pro-survival signals. Oncogene 2009; 28: 20742086.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 32.

    DeBoever S, Neirinkx E, Baert K, et al. Pharmacokinetics of tepoxalin and its active metabolite in broiler chickens. J Vet Pharmacol Therap 2008; 12: 97100.

    • Search Google Scholar
    • Export Citation
  • 33.

    Argentieri DC, Ritchie DM, Ferro MP, et al. Tepoxalin: a dual cyclooxygenase/5-lipoxygenase inhibitor of arachidonic acid metabolism and potent anti-inflammatory activity and a favorable gastrointestinal profile. J Pharmacol Exp Ther 1994; 271: 13991408.

    • Search Google Scholar
    • Export Citation
  • 34.

    Knight EV, Keenan CM, Smith IL, et al. Preclinical toxicity evaluation of tepoxalin, a dual inhibitor of cyclooxygenase and 5-lipoxygenase, in Sprague-Dawley rats and Beagle dogs. Fundam Appl Toxicol 1996; 33: 3848.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 35.

    Cicero AFG, Laghi L. Activity and potential role of licofelone in the management of osteoarthritis. Clin Interv Aging 2007; 2: 7379.

  • 36.

    Sundaram S, Ghosh J. Expression of 5-oxo-ETE receptor in prostate cancer cells: critical role in survival. Biochem Biophys Res Comm 2006; 339: 9398.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 37.

    Hayashi T, Nishiyama K, Shirahama T. Inhibition of 5-lipoxygenase pathway suppresses the growth of bladder cancer cells. Int J Urol 2006; 13: 10861091.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 38.

    Cianshi F, Cortesini C, Magnelli L, et al. Inhibition of 5-lipoxygenase by MK-886 augments the antitumor activity of celecoxib in human colon cancer cells. Mol Cancer Ther 2006; 5: 27162726.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 39.

    Fan XM, Tu SP, Lam SK, et al. Five-lipoxygenase activating protein inhibitor MK-886 induces apoptosis in gastric cancer through upregulation of p27kip1 and bax. J Gastroenterol Hepatol 2004; 19: 3137.

    • Crossref
    • Search Google Scholar
    • Export Citation
  • 40.

    Yoshida T, Shiraishi T, Horinaka M, et al. Lipoxygenase inhibitors induce death receptor5/Trail-R2 expression and sensitize malignant tumor cells to TRAIL-induced apoptosis. Cancer Sci 2006; 98: 14171423.

    • Search Google Scholar
    • Export Citation

Advertisement