59Fecl
3 were added to Ehrlich ascites cancer cell solution, the mixture were incubated in a water bath with shaking, and after certain times the uptakes of
59Fe by cancer cells, and effects of incubation temperature and of addition of carriers, metabolic inhibitors, amino acids, chelating agents, anti-cancer drugs and metal ions were investigated.
1) As for intracellular distribution of the taken up
59Fe, it was found in the greatest amount in the nuclear fraction, next in the supernatant and the microsornes. In the mitochondrial fraction, it appeared in the least amount.
2) Effects of incubation temperature : Between 16°C and 37°C, effects of temperature on the
59Fe uptake rate could scarcely be found.
3) By increasing cell number from 5×10
6 to 20×10
6 and 80×10
6, the
59Fe uptake rate increased, but the value per cell decreased contrarily.
4) On the addition of Fe
3+ (FeCl
3) as a carrier, the
59Fe uptake rate showed significant increase at 10
-3M, 10
-4M. On the addition of Fe
2+ (FeCl
2) as a carrier, the
59Fe uptake rate showed significant decrease at 20 minutes and increase at 60 minutes. When Fe
2+ and Fe
3+ were respectively added to 10
-6M, no significant changes could be found.
5) Effects of addition of metabolic inhibitors : The
59Fe uptake was remarkably inhibited at 10
-3M, of DNP, MIA and KCN, using 0.9%NaCl or Tris buffer as a medium, and at 10
-4 M-10
-5M the effect was not so much great. Only by addition of KCN to 10
-3M, using KRP as a medium, the uptake was contrarily promoted.
6) Effects of addition of various amino acid using 0.93/4NaCl as a medium were as follows : At 10
-3M, 10
-4M and 10
-5M of L-Cysteine, 10
-3M of L-Asparti acid and 10
-3M of Na Glutamate, the uptake showed significant increase. At 10
-3M of D-Penicillamine and L-Glutathione the significant decrease was observed. Addition of DL-Penicillamine, Glycine-Na and L-Histidine did not elicit any significant changes.
7) As for addition of various chelating agents using 0. 9%NaCl as a medium, the addition of Ca EDTA (10
-3M) and Dihydrothioctic acid (10
-3M) significantly decreased the uptake rate at 20 and 60 minutes. When Na citrate (10
-3M), Tetracycline (100μg), Ascorbic acid (10
-3M) and Riboflavin (200μg) were added however, the significant decrease was found only at 20 minutes, and when α-thiolactoyl glycine Na was added, no significant decrease could be found both at 20 and 60 minutes. The inhibition rate of
59Fe uptake rate by chelating agents including amino acid did not agree with the stability constant.
8) The effects of addition of various anti-cancer agents at various concentrations were as follows : 10μg of Toyomycin at 20 and 60 minutes, 100μg of Merphyrin at 60 minutes significantly decreased the uptake rate, however, 100μg of Nitromin at 20 and 60 : minutes conversely significantly increased it. As for the effects of Mitomycin C and Tespamin, any significant changes could not be found.
9) When various metal ions were added using 0.9%NaCl and Tris buffer as a medium, the degrees of uptake were varying without showing any regularity.
10) Addition of Na ATP, G-strophantin did not elicit any changes in
59Fe uptake rate.
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