Biological and molecular characterization of a canine hemangiosarcoma-derived cell line
Introduction
Altered endothelial cell proliferation and survival is of prime importance in many disease conditions, including neoplasia, wound healing, and chronic inflammatory disease. Perhaps the most dramatic example of dysregulated angiogenesis is seen in malignancy derived from vascular endothelium. Investigation of the mechanisms responsible for tumourigenesis in these neoplasms has the potential to shed light onto basic concepts of angiogenesis, and potentially identify novel strategies for its modulation.
Despite the fact that vascular tissue is extremely abundant, malignant tumours derived from the vascular endothelium are rare in humans, accounting for only 2% of soft-tissue sarcomas in one evaluation (Fata et al., 1999). When encountered, haemangiosarcomas of the human breast, liver, spleen and scalp behave aggressively, metastasizing commonly and yielding very poor long-term survival rates (Falk et al., 1979, Holden et al., 1987, Neuhauser et al., 2000, Silverman et al., 1994).
Haemangiosarcoma (HSA) is a relatively common, spontaneous tumour in dogs, accounting for approximately 20% of all soft-tissue sarcomas (Dorn et al., 1968), and up to 5% of all malignant canine neoplasms (Bastianello, 1983, MacVean et al., 1978). Canine HSA can occur in any site, however the spleen, skin, right atrium, and liver are the most common primary sites (MacEwen, 2001). Canine HSA is typified by very aggressive biological behavior, with wide and rapid metastasis being common. Doxorubicin-based chemotherapy has the potential to modestly improve survival time following surgery, however, one-year survival rates remain less than 10% (Sorenmo et al., 2000, Sorenmo et al., 1993, Vail et al., 1995).
Canine HSA has potential utility as a spontaneously occurring model of human endothelial tumours, such as angiosarcoma/hemangiosarcoma, hemangioma of infancy, and Kaposi’s sarcoma, a malignant tumour characterized by the dysregulated proliferation of endothelial cells. Moreover, canine HSA cells may also be useful as a tool for the study of endothelial cell biology in malignancy. For these purposes, we sought to establish a continuous cell line derived from a canine HSA, and to characterize this line with regard to its in vitro biological behavior, cell surface phenotype, and growth factor/growth factor receptor expression.
Section snippets
Cell lines
The DEN-HSA cell line was established from a spontaneous renal HSA arising in an 11-year-old castrated male golden retriever. The primary tumour was mechanically and enzymatically dissociated (collagenase III 50 U/mL, DNAse 100 U/mL, hyaluronidase 200 U/mL, Sigma, St. Louis, MO) (Kemmer et al., 1987), and passaged in tissue culture flasks in complete minimal essential medium (Cellgro Mediatech, Herndon, VA) supplemented with 5% fetal bovine serum, 5% newborn bovine serum, 100 U/mL penicillin, 100
Morphology
The DEN-HSA cell line has been morphologically stable in culture through >150 passages. When subconfluent, DEN-HSA cells appeared elongated and spindleoid, but adopted a more rounded, “cobblestone-like” morphology characteristic of EC from many species, including dogs (Gerhart et al., 1988) when confluent (Fig. 1(A)). Upon plating on Matrigel®, DEN-HSA formed microvascular tube-like structures (Fig. 1(B)). However, after 24 h these tube-like structures disappeared, and the cells appeared to form
Discussion
There are few endothelial-derived malignant tumour cell lines available for study. Those available include the Eoma murine endothelioma cell line (Obeso et al., 1990), and several artificially transformed murine endothelial cell lines, with in vivo characteristics similar to that of hemangioma or angiosarcoma (Arbiser et al., 2000, Arbiser et al., 1997, Montesano et al., 1990, RayChaudhury et al., 1994). More recently, cell line derived from spontaneous human and canine angiosarcoma have been
Acknowledgments
The authors dedicate this manuscript to their esteemed colleague and co-author, the late Dr. E. Gregory MacEwen, and to acknowledge the expert technical assistance of A.K. Marr, J.M. Schmidt, B. Charles and B. Shinners. This work was supported by the University of Wisconsin Animal Cancer Treatment Fund, and by a generous donation from Mr. A. Rolfe.
References (47)
- et al.
Interleukin-12 inhibits tumor growth in a novel angiogenesis canine hemangiosarcoma xenograft model
Neoplasia
(2004) - et al.
Overexpression of VEGF 121 in immortalized endothelial cells causes conversion to slowly growing angiosarcoma and high level expression of the VEGF receptors VEGFR-1 and VEGFR-2 in vivo
Am. J. Pathol.
(2000) - et al.
Hepatic angiosarcoma associated with androgenic-anabolic steroids
Lancet
(1979) - et al.
Immunohistochemical detection of CD31 antigen in normal and neoplastic canine endothelial cells
J. Comp. Pathol.
(1995) - et al.
Canine malignant hemangiosarcoma as a model of primitive angiogenic endothelium
Lab. Invest.
(2004) - et al.
Cultured human and canine endothelial cells from brain microvessels
Brain Res. Bull.
(1988) - et al.
Endotoxin and muramyl dipeptide modulate surface receptor expression on human mononuclear cells
Immunopharmacology
(2000) - et al.
Increased proteolytic activity is responsible for the aberrant morphogenetic behavior of endothelial cells expressing the middle T oncogene
Cell
(1990) - et al.
Splenic angiosarcoma: a clinicopathologic and immunophenotypic study of 28 cases
Mod. Pathol.
(2000) - et al.
Sickle cell anemia as a possible state of enhanced anti-apoptotic tone: survival effect of vascular endothelial growth factor on circulating and unanchored endothelial cells
Blood
(1999)
High urine concentrations of basic fibroblast growth factor in dogs with bladder cancer
J. Vet. Intern. Med.
Oncogenic H-ras stimulates tumor angiogenesis by two distinct pathways
Proc. Natl. Acad. Sci. USA
Monoclonal antibody against angiotensin-converting enzyme: Its use as a marker for murine, bovine, and human endothelial cells
Proc. Natl. Acad. Sci. USA
A survey on neoplasia in domestic species over a 40-year period from 1935 to 1974 in the republic of South Africa. VI. Tumors occurring in dogs
Onderspoort J. Vet. Res.
Biological roles of fibroblast growth factor-2
Endocr. Rev.
Expression of insulin-like growth factor receptor, IGF-1, and IGF-2 in primary and metastatic osteosarcoma
J. Surg. Oncol.
Plasma vascular endothelial growth factor concentrations in healthy dogs and dogs with hemangiosarcoma
J. Vet. Intern. Med.
Autocrine stimulation of VEGFR-2 activates human leukemic cell growth and migration
J. Clin. Invest.
Survey of animal neoplasms in Alameda and Contra Costa Counties. Cancer morbidity in dogs and cats from Alameda County
J. Natl. Cancer Inst.
Paclitaxel in the treatment of patients with angiosarcoma of the scalp or face
Cancer
Angiogenic activity of human soluble intercellular adhesion molecule-1
Cancer Res.
Expression of vascular endothelial growth factor and its receptor mRNA in angiosarcoma
Lab. Invest.
Expression of integrin alpha(v)beta(3) correlates with activation of membrane-type matrix metalloproteinase-1 (MT1-MMP) and matrix metalloproteinase-2 (MMP-2) in human melanoma cells in vitro and in vivo
Int. J. Cancer
Cited by (39)
Current understanding of comparative pathology and prospective research approaches for canine hemangiosarcoma
2024, Research in Veterinary ScienceImatinib and dasatinib inhibit hemangiosarcoma and implicate PDGFR-β and Src in tumor growth
2013, Translational OncologyFuture treatment of soft tissue sarcomas will be driven by histological subtype and molecular abberations
2010, European Journal of CancerCitation Excerpt :Furthermore, the selective mechanism of action for trabectedin in this translocation-related sarcoma is being studied, since the data suggest that specific changes in the sarcoma cells may sensitise to Trabectidin. Angiosarcomas are of endothelial origin, and their growth seems to be depending on the presence of angiogenic growth factor autocrine loops.9 While taxanes are considered to be inactive agents in soft tissue sarcomas, in general,10 they seem to yield activity against angiosarcomas.
Review of canine hemangiosarcoma: an aggressive and lethal neoplasm
2023, Veterinaria Mexico OAReview: The PI3K-AKT-mTOR signal transduction pathway in canine cancer
2023, Veterinary Pathology
- 1
Present address: Georgia Tech/IBB, 315 Forest Drive, Box 160, Atlanta, GA 30332, USA.
- 2
Present address: Primate Research Center, University of Wisconsin-Madison, 425 Henry Mall, Madison, WI 53706, USA.
- ✠
MacEwen is deceased.