Journal of Veterinary Clinics (한국임상수의학회지)

The Korean Society of Veterinary Clinics (한국임상수의학회)
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Establishment and Characterization of Canine Mammary Gland Cancer Cell Lines

개 자연발생 유선종양 2종의 세포주 확립 및 특성분석

  • Lee, Sun-Tae (Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University) ;
  • Kweon, Oh-Kyeong (Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University) ;
  • Kim, Wan-Hee (Department of Veterinary Surgery, College of Veterinary Medicine, Seoul National University)
  • Accepted : 2010.03.31
  • Published : 2010.06.30
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Abstract

Two cell lines derived from spontaneous canine mammary gland tumors were established and characterized. Mammary gland tumors from 9 years old pug and 9 years old toy-poodle dogs were collected by aseptic surgical resection and primary culture was performed. The histopathologic examination of tumors revealed adenocarcinoma and complex carcinoma and two dogs died from metastasis of the tumors. The tumor cells were subcultured over 60 times for more than 1 year and morphological consistency maintained. Light microscopic examination, growth curve, doubling time calculation, xenotransplantation to female nude mice, immunohistochemistry for wide spectrum keratin, vimentin, $\alpha$-smooth muscle actin and cytokeratin 8 was performed for characterization. The cell lines exhibited polygonal, elongated cell shape and cytoplasmic bridge and doubling time of 47.1 hrs and 18.6 hrs, respectively. Subcutaneous xenotransplantation to nude mice of the cells produced localized palpable mass within 4 weeks in 4 of 5 and 5 of 5 nude mice, respectively. In immunohistochemical examination one cell line showed strong positive against wide spectrum keratin and cytokeratin 8 and the other cell line showed strong positive against smooth muscle actin and cytokeratin 8. Additional characterization would be possible by investigator's needs and the cell lines may be useful for in vivo and in vitro studies of canine mammary tumor and adjuvant therapies.

개에서 자연발생한 유선종양으로부터 채취, 배양된 세포주 2개를 확립하고 특성을 분석하였다. 9세 령의 퍼그 종 개와 동일연령의 토이푸들 종의 개에 발생한 종양을 무균 상태를 유지하여 수술적으로 채취한 후 primary culture를 실시하였다. 두 종양의 조직병리검사에서는 각각 선암종과 혼합암종이 진단되었으며, 이후 두 마리 모두 전이로 인해 폐사하였다. 배양된 종양세포는 1년 이상의 기간 동안 60회 이상 계대를 반복하면서 형태학적으로 일관성을 유지하였고, 특성분석을 위해 광학현미경검사, 성장곡선 산출, 배가 시간 계산, 누드 마우스에 이종이식, 면역조직 화학검사를 실시하였다. 각 세포주는 다각형의 긴 세포형태를 보였으며, 세포질 연결을 형성하였으며, 배가 시간은 각각 47.1 시간과 18.6 시간이었다. 암컷 누드마우스의 등 부위에 피하이식 후 4주 이내에 10마리 중 9마리에서 촉진이 가능한 종괴의 형성이 확인되었으며, 면역조직화학검사 시 한 세포주에서는 keratin과 cytokeratin 8에서 다른 세포주에서는 smooth muscle actin과 cytokeratin 8에서 강한 염색성이 확인되었다. 두 세포주는 개의 유선종양의 시험관내, 체내 연구에 있어 모두 유용하게 사용될 수 있을 것으로 생각된다.

Keywords

References

  1. Brodey RS. Surgical treatment of cancer in the dog. J Am Vet Med Assoc 1975; 166: 494-496.
  2. Brower M, Carney DN, Oie HK, Gazdar AF, Minna JD. Growth of cell lines and clinical specimens of human nonsmall cell lung cancer in a serum-free defined medium. Cancer Res 1986; 46: 798-806.
  3. Campling BG, Haworth AC, Baker HM, Greer DL, Holden JJ, Bradley EC, Pym J, Dexter DF. Establishment and characterization of a panel of human lung cancer cell lines. Cancer 1992; 69: 2064-2074. https://doi.org/10.1002/1097-0142(19920415)69:8<2064::AID-CNCR2820690811>3.0.CO;2-#
  4. Destexhe E, Lespagnard L, Degeyter M, Heymann R, Coignoul F. Immunohistochemical identification of myoepithelial, epithelial, and connective tissue cells in canine mammary tumors. Vet Pathol 1993; 30: 146-154. https://doi.org/10.1177/030098589303000207
  5. Dorn CR, Taylor DO, Schenider R, Hibbard HH, Klauber MR. Survey of animal neoplasms in Alameda and Contra Costa Counties, California. II. Cancer morbidity in dogs and cats from Alameda County. J Natl Cancer Inst 1968; 40: 307-318.
  6. Fearon ER, Hamilton SR, Vogelstein B. Clonal analysis of human colorectal tumors. Science 1987; 238: 193-197. https://doi.org/10.1126/science.2889267
  7. Ferguson HR. Canine mammary gland tumors. Vet Clin North Am Small Anim Pract 1985; 15: 501-511. https://doi.org/10.1016/S0195-5616(85)50053-4
  8. Feuerhake F, Sigg W, Hofter EA, Dimpfl T, Welsch U. Immunohistochemical analysis of Bcl-2 and Bax expression in relation to cell turnover and epithelial differentiation markers in the non-lactating human mammary gland epithelium. Cell Tissue Res 2000; 299: 47-58. https://doi.org/10.1007/s004410050005
  9. Fialkow PJ. Clonal origin of human tumors. Biochim Biophys Acta 1976; 458: 283-321.
  10. Fidler IJ, Hart IR. The development of biological diversity and metastatic potential in malignant neoplasms. Oncodev Biol Med 1982; 4: 161-176.
  11. Fidler IJ, Naito S, Pathak S. Orthotopic implantation is essential for the selection, growth and metastasis of human renal cell cancer in nude mice [corrected]. Cancer Metastasis Rev 1990; 9: 149-165. https://doi.org/10.1007/BF00046341
  12. Gazdar AF, Oie HK. Re: Growth of cell lines and clinical specimens of human non-small cell lung cancer in a serumfree defined medium. Cancer Res 1986; 46: 6011-6012.
  13. Gilbertson SR, Kurzman ID, Zachrau RE, Hurvitz AI, Black MM. Canine mammary epithelial neoplasms: biologic implications of morphologic characteristics assessed in 232 dogs. Vet Pathol 1983; 20: 127-142. https://doi.org/10.1177/030098588302000201
  14. Hellmen E, Lindgren A. The expression of intermediate filaments in canine mammary glands and their tumors. Vet Pathol 1989; 26: 420-428. https://doi.org/10.1177/030098588902600507
  15. Heppner GH. Tumor heterogeneity. Cancer Res 1984; 44: 2259-2265.
  16. Nowell PC. Mechanisms of tumor progression. Cancer Res 1986; 46: 2203-2207.
  17. Perez-martinez C, Garcia-iglesias MJ, Duran-navarrete AJ, Espinosa-alvarez J, Garcia-fernandez RA, Lorenzana-robles N, Fernandez-perez S, Garcia-marin JF. Histopathological and immunohistochemical characteristics of two canine lipid-rich mammary carcinomas. J Vet Med A Physiol Pathol Clin Med 2005; 52: 61-66. https://doi.org/10.1111/j.1439-0442.2005.00690.x
  18. Rabanal RM, Else RW. Immunohistochemical localisation of cytokeratin and vimentin intermediate filament proteins in canine mammary tumours. Res Vet Sci 1994; 56: 225-233. https://doi.org/10.1016/0034-5288(94)90108-2
  19. Salsbury AJ. The significance of the circulating cancer cell. Cancer Treat Rev 1975; 2: 55-72. https://doi.org/10.1016/S0305-7372(75)80015-6
  20. Sartin EA, Barnes S, Toivio-kinnucan M, Wright JC, Wolfe LG. Heterogenic properties of clonal cell lines derived from canine mammary carcinomas and sensitivity to tamoxifen and doxorubicin. Anticancer Res 1993; 13: 229-236.
  21. Schneider R, Dorn CR, Taylor DO. Factors influencing canine mammary cancer development and postsurgical survival. J Natl Cancer Inst 1969; 43: 1249-1261.
  22. Sell S, Pierce GB. Maturation arrest of stem cell differentiation is a common pathway for thecellular origin of teratocarcinomas and epithelial cancers. Lab invest 1994; 70: 6-22.
  23. Sommers CL, Byers SW, Thompson EW, Torri JA, Gelmann EP. Differentiation state and invasiveness of human breast cancer cell lines. Breast Cancer Res Treat 1994; 31: 325-335. https://doi.org/10.1007/BF00666165
  24. Sorenmo KU, Shofer FS, Goldschmidt MH. Effect of spaying and timing of spaying on survival of dogs with mammary carcinoma. J Vet Intern Med 2000; 14: 266-270 https://doi.org/10.1111/j.1939-1676.2000.tb01165.x
  25. Taipale J, Beachy PA. The Hedgehog and Wnt signalling pathways in cancer. Nature 2001; 411: 349-354. https://doi.org/10.1038/35077219
  26. Zhou S, Schuetz JD, Bunting KD, Colapietro AM, Sampath J, Morris JJ, Lagutina I, Grosveld GC, Osawa M, Nakauchi H, Sorrentino BP. The ABC transporter Bcrp1/ABCG2 is expressed in a wide variety of stem cells and is a molecular determinant of the side-population phenotype. Nat Med 2001; 7: 1028-1034. https://doi.org/10.1038/nm0901-1028