Purification of canine alpha-fetoprotein and alpha- fetoprotein values in dogs

Yamada, Takatsugu; Kakinoki, Mika; Totsuka, Kazuya; Ashida, Yoshinori; Nishizono, Kazuya; Tsuchiya, Ryosuke; Kobayashi, Kosaku

doi:10.1016/0165-2427(94)05386-7

Cited by 14 publications

(17 citation statements)

References 28 publications

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“…Purification of human AFP Human AFP was prepared by the method as described elsewhere [9] . Briefly, human cord blood AFP was precipitated by ammonium sulfate and passed through an anti-AFP antibody affinity chromatography column.…”

Section: Methodsmentioning

confidence: 99%

Alpha-fetoprotein stimulated the expression of some oncogenes in human hepatocellular carcinoma Bel 7402 cells

Li¹,

Li²,

Chen³

et al. 2004

WJG

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AIM:To investigate the molecular mechanism of alphafetoprotein (AFP) on regulating the proliferation of human hepatocellular carcinoma cells. METHODS:Alpha-fetoprotein purified from human umbilical blood was added to cultured human hepatocellular carcinoma Bel 7402 cells in vitro for various treatment periods. The expression of c-fos, c-jun, and N-ras mRNA involved in proliferation and differentiation of cells was analyzed by Northern blot, and the expression of mutative p53 and p21 ras proteins was determined by Western blot. RESULTS:The results showed that AFP (20 mg/L) stimulated mRNA expression of these oncogenes in Bel 7402 cells. The expression of c-fos mRNA increased by 51.1%, 60.9%, 96.0%, and 25.5% at 2, 6, 12, and 24 h, respectively. The expression of c-jun and N-ras mRNA reached to the maximum which increased by 81.3% and 59.9% as compared with the control after 6 h and 24 h incubation with AFP, respectively. Western blot assay also demonstrated that AFP promoted the expression of mutative p53 and p21 ras proteins, and the increased rate of those proteins was 13.0%, 39.9%, and 70.9%, as well as 35.2%, 102.6%, and 46.8% at 6, 12, and 24 h, respectively, as compared with the control. Both human serum albumin (the same dosage as AFP) and monoclonal anti-AFP antibody failed to stimulate the expression of these oncogenes, but anti-AFP antibody could block the functions of AFP. CONCLUSION:The data indicate that AFP can stimulate the expression of some oncogenes to enhance the proliferation of human hepatocellular carcinoma Bel 7402 cells.Li MS, Li PF, Chen Q, Du GG, Li G. Alpha-fetoprotein stimulated the expression of some oncogenes in human hepatocellular carcinoma Bel 7402 cells. World J Gastroenterol 2004; 10(6): 819-824

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Section: Methodsmentioning

confidence: 99%

Alpha-fetoprotein stimulated the expression of some oncogenes in human hepatocellular carcinoma Bel 7402 cells

Li¹,

Li²,

Chen³

et al. 2004

WJG

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“…Careful examinations at the Veterinary Teaching Hospital of Azabu University revealed a high blood ALP activity (659 IU/L), and serum α-fetoprotein (AFP) level measured by ELISA methods [26] ranged from 400 to 500 ng/ml during the next 2 years after the first admission.…”

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Hepatoblastoma in a Dog.

Shiga

Shirota

Shida

et al. 1997

The Journal of Veterinary Medical Science

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ABSTRACT. A hepatoblastoma was found in a 13-year-old female Maltese dog. Histologically, the tumor showed a wide trabecular pattern and was frequently accompanied with vascular lake formation. Tumor cells were positive for cytokeratin and neuron specific enolase, but negative for chromogranin. Electronmicroscopically, tumor cells were accompanied with continuous basement membrane and had poorly developed desmosomes. Sinusoidal endothelia had fenestration and were surrounded by myofibroblast-like cells. To the best of our knowledge, this paper is the first report of morphological studies on canine hepatoblastoma. -KEY WORDS: canine, hepatoblastoma.J. Vet. Med. Sci. 59(12): 1167-1170, 1997 periodic acid-Schiff (PAS) reaction with or without diastase digestion, Orcein stain, and Watanabe's silver impregnation.Mitoses of tumor cells were counted and the data were compared with those of a canine hepatocellular carcinoma with distant metastasis.Immunohistochemical examination was performed using labelled streptavidin-biotin method with a Histofine kit (Nichirei Corp., Tokyo, Japan). Primary antibodies used were as follows: anti-human cytokeratin antibody (Boehringer Mannheim Biochemica, Mannheim, Germany), anti-human neuron specific enolase (NSE) antibody (Dakopatts, Glostrup, Denmark), anti-bovine chromogranin antibody (Incstar Corp., Stillwater, MN, U.S.A.), anti-α-smooth muscle actin (α-SMA) antibody (Sigma Chemical Co., St. Louis, MO, U.S.A.), and anti-human factor VIIIrelated antigen (FVIII-RAg) antibody (Dakopatts, Glostrup, Denmark). Immunoreaction with primary antibodies was performed for 1 hr at 37°C. Thereafter, biotinylated antirabbit or mouse IgG antibody and peroxidase-labelled streptavidin were sequentially applied to the sections for 15 min each at room temperature. The reaction was visualized with 3,3'-diaminobenzidine in 0.1M Tris HCl buffer (pH 7.6) plus hydrogen peroxide. Sections were counterstained with Mayer's hematoxylin.For electron microscopy, small blocks (1 mm 3 ) of the tumor were fixed in 2.5% glutaraldehyde/0.1 M phosphate buffer for 2 hr at 4°C and post-fixed in 1% osmium tetroxide/0.2 M phosphate buffer for 2 hr at 4°C. The blocks were dehydrated in graded ethanol and embedded in Epoxy resin. Ultra-thin sections were cut and double stained with uranyl acetate and lead citrate and examined by a Hitachi H-300 transmission electron microscope at 75 kV.Histologically, the nodules were consisted of compact proliferation of tumor cells in a wide trabecular pattern (Fig. 1). Many vascular lakes were present in the tumor tissues (Fig. 1). Necrosis and degeneration were rare. However, in the central portions of tumor nests, neoplastic cells fell into degeneration (Fig. 2a) and these portions occasionally connected to the vessels in the stroma (Fig. 2b). The tumor cells had a polarity: their nuclei were located at the opposite site of basal side (Fig. 2a). The nuclei of the tumor cells were round with relatively rich chromatin and did not show atypia or pleomorphism. The nucleus/cytoplasm ratio...

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“…Yamada et al previously developed an ELISA for canine AFP to investigate changes in serum AFP in newborn puppies. 13 Subsequently, they found that, as in humans, AFP concentrations are elevated in canine tumors, especially in hepatocellular carcinoma (HCC). 12 In those reports, serum AFP was mildly elevated in some dogs with hepatic and nonhepatic diseases, even in those without tumors.…”

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confidence: 99%

“…g The tissue section was stained using an avidin-biotin complex method with biotinylated rabbit antidog AFP IgG, 13 peroxidase-conjugated streptavidin g and diaminobenzidine.…”

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confidence: 99%

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Alpha-Fetoprotein in Serum and Tumor Tissues in Dogs with Hepatocellular Carcinoma

Kitao¹,

Yamada²,

Ishikawa³

et al. 2006

J VET Diagn Invest

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Abstract. Serum alpha-fetoprotein (AFP) concentrations were measured before and after surgical removal of tumor masses in four dogs with hepatocellular carcinoma (HCC). Localization of AFP was also examined immunohistochemically in tumor tissues. In three cases, the serum AFP concentration was 10 to 20 times higher than that of normal dogs. One to two months after surgery, the serum AFP concentration had decreased to normal range. AFP was localized in the tumor tissues in these three cases. One case, which had a low serum AFP, did not show AFP localization in tumor tissue.

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Purification of canine alpha-fetoprotein and alpha- fetoprotein values in dogs

Cited by 14 publications

References 28 publications

Alpha-fetoprotein stimulated the expression of some oncogenes in human hepatocellular carcinoma Bel 7402 cells

Alpha-fetoprotein stimulated the expression of some oncogenes in human hepatocellular carcinoma Bel 7402 cells

Hepatoblastoma in a Dog.

Alpha-Fetoprotein in Serum and Tumor Tissues in Dogs with Hepatocellular Carcinoma

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