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PubMed1993

Lack of copper binding sites in ceruloplasmin of LEC rats with abnormal copper metabolism.

Recently it was found that the clinical features of the LEC rat closely resemble those of human Wilson's disease. One of the characteristics of the animal is low levels of serum ceruloplasmin. Therefore, by using LEC rats, we attempted to define molecular basis of the deficiency in active site of ceruloplasmin in Wilson's disease patients. We made 3 monoclonal antibodies, ID2 against active site of ceruloplasmin, ID1 against inactive site of ceruloplasmin, and the remaining one against metallothionein. Using these monoclonal antibodies, we examined immunohistochemical stainings of LEC rat liver tissues, and compared them with those of LEA rats, as a control. ID1 stained the hepatocytes of both LEA and LEC rats, whereas ID2 stained LEA rat hepatocytes only. The results indicated that the ceruloplasmin secreted by LEC rat hepatocytes is mostly in inactive form. The antibody against metallothionein stained LEA rat hepatocytes only. This finding may also indicate that LEC rat hepatocytes express less amount of metallothionein than those of LEA rats.

Animals↗
PubMed1993

Suppression of 9L gliosarcoma growth by copper depletion with copper-deficient diet and D-penicillamine.

Trace element such as Cu and Zn have important chemical and biological properties. Recently, tissue Cu and Zn concentrations have been correlated with prognosis in selected malignancies. In addition, depletion of trace metals has suppressive effects on tumor growth in experimental rat models. We measured tissue levels of Cu and Zn and investigated the inhibitory effects on tumor growth in a rat brain tumor model by the Cu-depletion. 9L gliosarcoma cells were injected subcutaneously in 24 anesthetized 5 week old male Fischer-344 rats. Control animals (n = 12) were given a normal diet throughout the experiment and hypocupremic rats (n = 12) were given a Cu-deficient diet beginning 3 weeks before and after tumor implantation, and administered 2 mg of D-Penicillamine peri os., once daily, 3 days before and after implantation. At the time of sacrifice, samples were taken to measure tumor weights. To determine tissue Cu and Zn levels, atomic absorption spectrophotometry was used. Cu, Zn and Cu/Zn ratio were significantly higher in control tumors than brain tissues. Cu levels and Cu/Zn ratio were significantly lower in hypocupremic tumors than those in control tumors. Zn levels in hypocupremic tumors were significantly higher than control tumors. Our study indicated that Cu depletion by a Cu-deficient and D-Penicillamine reduced Cu concentration and Cu/Zn ratio in a tumor model with reduction of tumor weight. A metabolic approach that restricts Cu to alter the microenvironment within the cell must become a new horizon of cancer therapy.

Animals↗