- Email: [email protected]
Mouse liver free amino acids during the development of Ehrlich ascites tumour
Cancer
Letters.
58 (1991)
22
221-224
Elsevier Scientific Publishers
Ireland
Ltd.
Mouse liver free amino acids during the development ascites tumour J. Mdrquez Departomento
of Ehrlich
and I. Niiiiez de Castro de Bioquimica
y Biologh Molecular, Facultad de Ciencias, Uniuersidod de M6laga (Spain)
(Received 22 January 1991) (Revision received 17 April 1991) (Accepted 1 May 1991)
Summary
Introduction
Sequential amino acid concenfrations were determined in the lioer of mice infested with a highly malignant strain of Ehrlich ascites tumour cells. The liuer concentrations of a certain group of amino acids showed changes
Tumour growth elicits systemic metabolic changes in the host resulting from the tumour consumption of nutrients and from the output of certain unknown biochemical sighals [Ml. Kawamura et al. [5] reported significant alterations in liver weight and protein metabolism in advanced stages of experimental neoplastic disease. This enlargement of liver was also observed by Tessitore et al. [16] in the first days after tumour transplantation associated with an increment of protein content; in the same way, an early impairment of host liver mitochondria has been detected [17]. An imbalanced ornithine metabolism in liver of mice bearing Ehrlich ascites carcinoma have been detected associated with tumour demand for polyamines [lo]. It is well established that variations of free amino acids in the host-tumour system during tumour development reveals a continous interplay between the rates of amino acid production and consumption by both the tumour and host tissues [l l] . Glutamine appears to be the main nitrogen source for rapidly dividing cells [1,6,23]. A net flux of glutamine from the host to the tumour was confirmed in vivo in mice inoculated with Ehrlich ascites tumour cells [2]; therefore, the whole organism is mobilized to augment the circulating glutamine 1141. The present study was carried out to
consistent
with
those
preoiously
reported
for
plasma, ascitic liquid and tumour cells during tumour growth. Shortly after tumour transplantation a significant decrease of the essential amino acids methionine, threonine, ualine, isoleucine + phenylalanine, leucine, lysine and histidine, was detected. Some nonessential amino acids, mainly the gluconeogenic substrates alanine and serine, showed a strong reduction in hepatic concentrations during the first days; these amino acids remained significantly lower than controls until animal death. Interestingly, hepatic glutamine increased at days 1 and 2 after inoculation, and proline showed a sustained increase from the seventh day onwards, reaching a ualue double
the control
Keywords: tumour
at the end of animal
life.
amino acids; liver; Ehrlich ascites
Correspondence to: I. NGiiez de Castro, Department0 de Bioquimica y Biologfa Molecular, Facultad de Ciencias, Universidad de MBlaga. 29071 Mdlaga, Spain.
0304-3835/91/$03.50 Published and Printed
0 1991 Elsevier Scientific Publishers in Ireland
Ireland
Ltd
222
determine the sequential changes of liver free amino acids over the whole period of tumour development. Materials Ehrlich
and Methods
ascites
cells
A hyperdiploid Lettre strain was maintained in Z-month-old female albino Swiss mice OF1 (SPF Ice), as described elsewhere [12]. The life span of the animals after inoculation with 5 x lo6 tumour cells was 16 f 1 days. Preparation
of samples
For amino acid determinations in liver, several groups of mice were inoculated with 5 x lo6 tumour cells from at least 4 different infested animals and sequential analyses of the whole series were carried out 1, 2, 4, 7, and 14 days after tumour transplantation. Four nontransplanted animals were used as controls. The animals were sacrificed by cervical dislocation; the liver was immediatety (less than 15 s) excised and frozen in liquid nitrogen, in a precooled mortar. It was then weighed, triturated, treated with four volumes of cold 1 M HC104 and centrifuged at 2000 x g for 5 min. The supernatants were neutralized with cold KOH solution, centrifuged again and kept at - 30°C until analysis. Amino
acid
analysis
The free amino acid concentrations were determined by a high-performance liquid chromatographic method fully validated for using precolumn dansyl biological samples, derivatization [9]. Water content of wet liver was assumed to be 71.6% (w/w) [21]. Values are expressed as means f S.E.; Student’s ttest for statistical significance was used. Results and Discussion Even though the interpretation of the hepatic free amino acid profiles is obscured due to the multiple metabolic pathways altered by the presence of a tumour, for specific groups of amino acids several analogous pat-
terns emerge. The sequential changes in the hepatic non-essential amino acid concentrations are shown in Table I. Liver glutamine displayed a very similar pattern to that observed in plasma [ 111, accounting for the simultaneous regulation of host tissues glutaminase and glutamine synthetase activities, which enable a net production of glutamine by the host during the initial days of tumour development 1141. Pain and Garlick [13] have reported a 40% increase in the hepatic protein synthesis in mice bearing Ehrlich ascitic carcinoma. There is also an accelerated protein synthesis in Morris hepatomas with a significant enhanced incorporation of aspartate and alanine in the tumour protein [19]. Such evidence could support the decrease in the concentrations of aspartate, alanine, serine and most of the essential amino acids (Table II) including threonine, valine, isoleucine, leucine, phenylalanine and methionine, during the first days of tumour growth. These results parallel with plasma concentrations of these essential amino acids, which decreased after tumour transplantation and only returned to control values at the final days of life, when tumour proliferation has already ceased [ 111. On the other hand, there have been several reports showing that an increased gluconeogenesis is elicited in the host by the presence of tumour both in humans [7] and tumour-bearing animals [15]. Fifty percent of the hepatic glucose comes from alanine [3]; a decrease in liver alanine content has been detected in gluconeogenic rats [22], and enhanced gluconeogenesis from alanine is reported in cancer patients [20]. From the fifth day onwards, plasma glucose levels decrease in mice bearing Ehrlich ascites tumour [15]. There is also an additional hypoglucemic pressure originated by a decreased food intake between the 8 and 12 days following inoculation [8]. It is notewhorty that alanine, serine and threonine concentrations remained lower than controls during almost the whole period of tumour development. The decreased availability in liver of these amino acids could
223
Table 1. Liver concentrations Amino acid
GIY
Ala Gln Glu Asn ASP
Ser Pro Arg Tyr CYS
Tau
(mM) of non-essential
Days after tumour
transplantation
0
1
1.72 1.11 2.08 1.40 0.04 0.18 0.35 0.14 0.08 0.03 0.67 2.91
* f zt f f + f zt zt f + f
0.04 0.1 0.2 0.09 0.004 0.01 0.01 0.01 0.01 0.001 0.06 0.21
1.91 0.55 3.36 1.70 0.05 0.09 0.25 0.15 0.08 0.03 0.91 2.65
amino acids of tumour bearing mice during tumour development.
f f zt f f f zt f zt f f f
0.04 0.03b 0.2” 0.07 0.005 0.01” O.Olb 0.01 0.003 0.001 0.07 0.05
1.56 0.19 2.72 0.67 0.05 0.09 0.23 0.09 0.06 0.01 1.14 2.36
f f f zt f f + f zt f f f
Values are means of at least 4 livers of different infested “P < 0.001; bP <0.005; dP <0.02. versus the control
animals value.
reflect the enhanced hepatic gluconeogenesis elicited by the tumour. The hepatic proline concentrations showed a continuous increase from the seventh day on, reaching a value double as control at the end of the animal life. A concentration gradient of proline from the ascitic fluid towards the plasma was established in mice bearing Ehrlich ascites carcinoma, with proline plasma concentrations three times higher than controls Table 11. Liver Amino acid
2.33 0.96 2.70 1.11 0.05 0.17 0.16 0.14 0.07 0.03 1.15 2.50
0.16 0.02” 0.12d 0.04” 0.003 0.01” 0.01” 0.004 0.002 0.001” 0.02” 0.11
14
7
4
2
f 0.08” * 0.08 + 0.2 f 0.03 f 0.001 f 0.03 ZJZ0.02” f 0.01 f 0.001 f 0.001 f 0.03” f 0.06
1.80 0.72 2.27 1.56 0.05 0.14 0.19 0.24 0.07 0.03 1.28 2.61
+ +z f zt f f f zt f f + +
0.2 0.06” 0.3 0.2 0.007 0.03 0.01” 0.03d 0.01 0.01 0.04” 0.04
1.46 0.69 1.51 0.67 0.05 0.11 0.15 0 31 0.03 0.02 1.12 2.33
+ +z f + l
f f * f f f zt
0.2 0.06” 0.15 0.1” 0.003 0.02 0.01” 0.03” 0.003” 0.001 0.02” 0.01
* S.E.
at the eleventh day [ll]. These findings strengthen the possibility of an impairment of liver to metabolize the enhanced proline
SUPPlY. Concerning the sulphur amino acids, methionine concentrations remained significantly lower than controls during the exponential phase of tumour growth. Ehrlich ascites cells maintain a very active polyamine biosynthesis [4] and the host ornithine metabo-
concentrations (mM) of essential amino acids of tumour bearing mice during tumour development.
Days after tumour
transplantation
0
1
Met Thr Val Ile + Phe Leu
0.09 0.16 0.29
zt 0.06 f 0.003 zt 0.01
0.08 0.12 0.24
0.11 0.12
f f
0.01 0.01
LYS His
0.24 0.55
+ 0.01 * 0.02
2
14
7
4
f f f
0.004 O.Olb 0.01
0.06 0.11 0.17
zt 0.003” f O.Old f 0.01”
0.05 0.11 0.17
+ 0.01’ f O.Olb f 0.01”
0.07 0.15 0.15
f 0.01 f 0.004 f 0.01”
0.08 zt 0.01 0.11 f 0.01 0 19 f 0.01
0.09
f
0.004
0.08
f
0.003b
0.07 0.06
0.24 0.33
ziz 0.02 f 0.02”
it 0.003’ f O.Old f 0.02 f 0.01
0.08 0.09 0.16 0.52
i f * zt
0.08 0.05 0.18 0.47
f 0.01 f O.Olb zt 0.02 l 0.02
0.11 0.11 0.23 0.65
0.22 0.57
0.01 0.01 0.01’ 0.02
Values are means of at least 4 livers of different infested animals f S.E. “P < 0.001; bP < 0.005; ‘P < 0.01; dP < 0.02, versus the control value.
f 0.01 f 0.01 ziz 0.02 zt 0.03
224
lism seems to be adapted to a net production of this amino acid needed for tumour growth [lo]. One of the main metabolic fates of methionine is polyamine biosynthesis, so the diminished hepatic contents of this amino acid may partially reflect its preferential shift toward tumour polyamines, paralleling the behaviour of ornithine. Liver cysteine increased in tumour bearing mice along tumour development; the same pattern was observed in plasma [ 111. The results presented here confirm that after tumour transplantation the nitrogen metabolism of the host liver is altered reflecting changes in the free amino acid pools along the tumour development. Acknowledgement This study was supported PB/88/0455 from the Direcci6n InvestigacSn Cientifica y Tknica.
9
10
11
12
13
14
by Grant General de
713-719. Carrascosa, J.M., Martinez, P. and N6tiez de Castro, I. (1984) Nitrogen movement between host and tumor in mice inoculated with Ehrlich ascitic tumor cells. Cancer Res., 44, 3831-3835. Felig, P.. Pozefsky, T., Marliss, E. and Cahill, (1970) Alanine: key role in gluconeogenesis. 167, 1003-4.
G.F., Jr. Science,
Janne, J., Poso, H. and Raina. A. (1978) Polyamines in rapid growth and cancer. Biochim. Biophys. Acta, 473, 241-293. Kawamura. I., Moldawer, L.L., Keenan, R.A., Batist. G., Bothe, A., Jr., Altered amino growth. Cancer Laze, P. (1981)
Quesada. A.R. and NGfiez de Castro, 1. (1989) Nitrogen metabolism in tumor bearing mice. Arch. Biochem. Biophys., 268, 667-675. Olavarria. J.S., Chico, E., GimBnez-Gallego, G. and NdRez de Castro, I. (1981) Effect of ammonium ions on the aerobic glycolysis in Ehrlich ascites tumor cells. Biochimie. 63. 469-475. Pain, V.M. and Garlick. P.J. (1980) The effect of an Ehrlich ascites tumour on the rate of protein synthesis in muscle and liver of the host. Biochem. Sot. Trans., 8,354. Quesada, A.R.. Medina. M.A., Mbrquez, J.. SbnchezJimenez. F. and Ndtiez de Castro, 1. (1988) Contribution
15
by host tissues to circulating glutamine in mice inoculated with Ehrlich ascites tumor ceils. Cancer Res.. 48. 1551-1553. Shapot, V.S. (1980) Manifestations and mechanisms of
16
systemic effect of tumours on the host. In: Biochemical aspects of tumour growth. pp. 124-173. Editor: D.A. Myshne. MIR Publishers, Moscow. Tessitore. L., Bonelli, G. and Baccino. F.M. (1987) Early
References Ardawi. M.S.M. and Newsholme, E.A.(1985) Fuel utilization in colonocytes of the rat. Biochem. J.. 231.
M6rquez, F.J., Quesada, A.R.. Sbnchez-JimPnez, F. and N6fiez de Castro, I. (1986) Determination of 27 Dansyl amino acid derivatives in biological fluids by reversedphase high performance liquid chromatography. J. Chromatogr., 380. 275-283. MBrquez, J., M&es. J .M., Quesada, A.R., Medina, M .A., N(liiez de Castro, I. and SBnchez-JimQnez, F. (1989) Altered ornithine metabolism in tumor-bearing mice. Life Sci., 45, 1877-1884. MBrquez. J., S6nchez-JimLnez, F., Medina. M.A.,
17
18
19
development of protein metabolic perturbations in the liver and skeletal muscle of tumour-bearing rats. Biochem. J.. 241, 153-159. Urdiales. J.L., Medina, M.A., Ndtiez de Castro, I. and SBnchez-Jimhnez, F. (1989) Early systemic effects on the hepatic mitochondria of tumour bearing mice. Cancer Lett.. 44, 179-183. Vincent, M.D. (1985) The clinical problem. In: The molecular basis of cancer, pp. l-35. Editors: P.B. Farmer and J.M. Walker. Croom Helm, London. Wagle, S.R., Morris. H.P. and Weber, G. (1963) Comparative biochemistry of hepatomas. V. Cancer Res., 23. 1003- 1007. Waterhouse, C., Jeanpretre, N. and Keilson, J. (1979) Gluconeogenesis from alanine in patients with progressive malignant disease. Cancer Res.. 39. 1968-1972.
Bistrian, B.R. and Blackburn. G.L. (1982) acid kinetics in rats with progressive tumor Res., 42, 824-829. Amino acids and glucose utilization by dif-
21
ferent metabolic pathways in ascites tumour cells. Eur. J. Biochem., 117, 1925. Levin. L., Gevers. W., Jardine, L., De Guel, F.J.M. and Duncan, E.J. (1983) Serum amino acids in weight-losing
West, J.T. book W.R.
22
patients with cancer and tuberculosis. Eur. J. Cancer Clin. Oncol., 19, 711-715. MBrquez, F.J. (1987) Estudio din6mico de amino6cidos libres en el sistema huesped-tumor en ratones inoculados
Williamson, D.H.. Lopes-Vieira. 0. and Walker, B. (1967) Concentrations of free glucogenic amino acids in livers of rats subjected to various detabolic stresses. Biochem. J.. 104. 497-502.
23
Windmueller. H.G. and Spaeth. metabolism of plasma glutamine Biol. Chem., 249. 5070-5079.
con el carcinoma sity of Mhlaga.
ascitico de Ehrlich. Ph.D. Thesis, Univer-
20
ES., Todd, W.R., Mason, H.S. and Van Bruggen, (1966) General composition of the body. In: Textof Biochemistry, pp. 416. Editors: E.S. West and Todd. The Macmillan Company, New York.
A.E. (1974) Uptake and by the small intestine. J.
Letters.
58 (1991)
22
221-224
Elsevier Scientific Publishers
Ireland
Ltd.
Mouse liver free amino acids during the development ascites tumour J. Mdrquez Departomento
of Ehrlich
and I. Niiiiez de Castro de Bioquimica
y Biologh Molecular, Facultad de Ciencias, Uniuersidod de M6laga (Spain)
(Received 22 January 1991) (Revision received 17 April 1991) (Accepted 1 May 1991)
Summary
Introduction
Sequential amino acid concenfrations were determined in the lioer of mice infested with a highly malignant strain of Ehrlich ascites tumour cells. The liuer concentrations of a certain group of amino acids showed changes
Tumour growth elicits systemic metabolic changes in the host resulting from the tumour consumption of nutrients and from the output of certain unknown biochemical sighals [Ml. Kawamura et al. [5] reported significant alterations in liver weight and protein metabolism in advanced stages of experimental neoplastic disease. This enlargement of liver was also observed by Tessitore et al. [16] in the first days after tumour transplantation associated with an increment of protein content; in the same way, an early impairment of host liver mitochondria has been detected [17]. An imbalanced ornithine metabolism in liver of mice bearing Ehrlich ascites carcinoma have been detected associated with tumour demand for polyamines [lo]. It is well established that variations of free amino acids in the host-tumour system during tumour development reveals a continous interplay between the rates of amino acid production and consumption by both the tumour and host tissues [l l] . Glutamine appears to be the main nitrogen source for rapidly dividing cells [1,6,23]. A net flux of glutamine from the host to the tumour was confirmed in vivo in mice inoculated with Ehrlich ascites tumour cells [2]; therefore, the whole organism is mobilized to augment the circulating glutamine 1141. The present study was carried out to
consistent
with
those
preoiously
reported
for
plasma, ascitic liquid and tumour cells during tumour growth. Shortly after tumour transplantation a significant decrease of the essential amino acids methionine, threonine, ualine, isoleucine + phenylalanine, leucine, lysine and histidine, was detected. Some nonessential amino acids, mainly the gluconeogenic substrates alanine and serine, showed a strong reduction in hepatic concentrations during the first days; these amino acids remained significantly lower than controls until animal death. Interestingly, hepatic glutamine increased at days 1 and 2 after inoculation, and proline showed a sustained increase from the seventh day onwards, reaching a ualue double
the control
Keywords: tumour
at the end of animal
life.
amino acids; liver; Ehrlich ascites
Correspondence to: I. NGiiez de Castro, Department0 de Bioquimica y Biologfa Molecular, Facultad de Ciencias, Universidad de MBlaga. 29071 Mdlaga, Spain.
0304-3835/91/$03.50 Published and Printed
0 1991 Elsevier Scientific Publishers in Ireland
Ireland
Ltd
222
determine the sequential changes of liver free amino acids over the whole period of tumour development. Materials Ehrlich
and Methods
ascites
cells
A hyperdiploid Lettre strain was maintained in Z-month-old female albino Swiss mice OF1 (SPF Ice), as described elsewhere [12]. The life span of the animals after inoculation with 5 x lo6 tumour cells was 16 f 1 days. Preparation
of samples
For amino acid determinations in liver, several groups of mice were inoculated with 5 x lo6 tumour cells from at least 4 different infested animals and sequential analyses of the whole series were carried out 1, 2, 4, 7, and 14 days after tumour transplantation. Four nontransplanted animals were used as controls. The animals were sacrificed by cervical dislocation; the liver was immediatety (less than 15 s) excised and frozen in liquid nitrogen, in a precooled mortar. It was then weighed, triturated, treated with four volumes of cold 1 M HC104 and centrifuged at 2000 x g for 5 min. The supernatants were neutralized with cold KOH solution, centrifuged again and kept at - 30°C until analysis. Amino
acid
analysis
The free amino acid concentrations were determined by a high-performance liquid chromatographic method fully validated for using precolumn dansyl biological samples, derivatization [9]. Water content of wet liver was assumed to be 71.6% (w/w) [21]. Values are expressed as means f S.E.; Student’s ttest for statistical significance was used. Results and Discussion Even though the interpretation of the hepatic free amino acid profiles is obscured due to the multiple metabolic pathways altered by the presence of a tumour, for specific groups of amino acids several analogous pat-
terns emerge. The sequential changes in the hepatic non-essential amino acid concentrations are shown in Table I. Liver glutamine displayed a very similar pattern to that observed in plasma [ 111, accounting for the simultaneous regulation of host tissues glutaminase and glutamine synthetase activities, which enable a net production of glutamine by the host during the initial days of tumour development 1141. Pain and Garlick [13] have reported a 40% increase in the hepatic protein synthesis in mice bearing Ehrlich ascitic carcinoma. There is also an accelerated protein synthesis in Morris hepatomas with a significant enhanced incorporation of aspartate and alanine in the tumour protein [19]. Such evidence could support the decrease in the concentrations of aspartate, alanine, serine and most of the essential amino acids (Table II) including threonine, valine, isoleucine, leucine, phenylalanine and methionine, during the first days of tumour growth. These results parallel with plasma concentrations of these essential amino acids, which decreased after tumour transplantation and only returned to control values at the final days of life, when tumour proliferation has already ceased [ 111. On the other hand, there have been several reports showing that an increased gluconeogenesis is elicited in the host by the presence of tumour both in humans [7] and tumour-bearing animals [15]. Fifty percent of the hepatic glucose comes from alanine [3]; a decrease in liver alanine content has been detected in gluconeogenic rats [22], and enhanced gluconeogenesis from alanine is reported in cancer patients [20]. From the fifth day onwards, plasma glucose levels decrease in mice bearing Ehrlich ascites tumour [15]. There is also an additional hypoglucemic pressure originated by a decreased food intake between the 8 and 12 days following inoculation [8]. It is notewhorty that alanine, serine and threonine concentrations remained lower than controls during almost the whole period of tumour development. The decreased availability in liver of these amino acids could
223
Table 1. Liver concentrations Amino acid
GIY
Ala Gln Glu Asn ASP
Ser Pro Arg Tyr CYS
Tau
(mM) of non-essential
Days after tumour
transplantation
0
1
1.72 1.11 2.08 1.40 0.04 0.18 0.35 0.14 0.08 0.03 0.67 2.91
* f zt f f + f zt zt f + f
0.04 0.1 0.2 0.09 0.004 0.01 0.01 0.01 0.01 0.001 0.06 0.21
1.91 0.55 3.36 1.70 0.05 0.09 0.25 0.15 0.08 0.03 0.91 2.65
amino acids of tumour bearing mice during tumour development.
f f zt f f f zt f zt f f f
0.04 0.03b 0.2” 0.07 0.005 0.01” O.Olb 0.01 0.003 0.001 0.07 0.05
1.56 0.19 2.72 0.67 0.05 0.09 0.23 0.09 0.06 0.01 1.14 2.36
f f f zt f f + f zt f f f
Values are means of at least 4 livers of different infested “P < 0.001; bP <0.005; dP <0.02. versus the control
animals value.
reflect the enhanced hepatic gluconeogenesis elicited by the tumour. The hepatic proline concentrations showed a continuous increase from the seventh day on, reaching a value double as control at the end of the animal life. A concentration gradient of proline from the ascitic fluid towards the plasma was established in mice bearing Ehrlich ascites carcinoma, with proline plasma concentrations three times higher than controls Table 11. Liver Amino acid
2.33 0.96 2.70 1.11 0.05 0.17 0.16 0.14 0.07 0.03 1.15 2.50
0.16 0.02” 0.12d 0.04” 0.003 0.01” 0.01” 0.004 0.002 0.001” 0.02” 0.11
14
7
4
2
f 0.08” * 0.08 + 0.2 f 0.03 f 0.001 f 0.03 ZJZ0.02” f 0.01 f 0.001 f 0.001 f 0.03” f 0.06
1.80 0.72 2.27 1.56 0.05 0.14 0.19 0.24 0.07 0.03 1.28 2.61
+ +z f zt f f f zt f f + +
0.2 0.06” 0.3 0.2 0.007 0.03 0.01” 0.03d 0.01 0.01 0.04” 0.04
1.46 0.69 1.51 0.67 0.05 0.11 0.15 0 31 0.03 0.02 1.12 2.33
+ +z f + l
f f * f f f zt
0.2 0.06” 0.15 0.1” 0.003 0.02 0.01” 0.03” 0.003” 0.001 0.02” 0.01
* S.E.
at the eleventh day [ll]. These findings strengthen the possibility of an impairment of liver to metabolize the enhanced proline
SUPPlY. Concerning the sulphur amino acids, methionine concentrations remained significantly lower than controls during the exponential phase of tumour growth. Ehrlich ascites cells maintain a very active polyamine biosynthesis [4] and the host ornithine metabo-
concentrations (mM) of essential amino acids of tumour bearing mice during tumour development.
Days after tumour
transplantation
0
1
Met Thr Val Ile + Phe Leu
0.09 0.16 0.29
zt 0.06 f 0.003 zt 0.01
0.08 0.12 0.24
0.11 0.12
f f
0.01 0.01
LYS His
0.24 0.55
+ 0.01 * 0.02
2
14
7
4
f f f
0.004 O.Olb 0.01
0.06 0.11 0.17
zt 0.003” f O.Old f 0.01”
0.05 0.11 0.17
+ 0.01’ f O.Olb f 0.01”
0.07 0.15 0.15
f 0.01 f 0.004 f 0.01”
0.08 zt 0.01 0.11 f 0.01 0 19 f 0.01
0.09
f
0.004
0.08
f
0.003b
0.07 0.06
0.24 0.33
ziz 0.02 f 0.02”
it 0.003’ f O.Old f 0.02 f 0.01
0.08 0.09 0.16 0.52
i f * zt
0.08 0.05 0.18 0.47
f 0.01 f O.Olb zt 0.02 l 0.02
0.11 0.11 0.23 0.65
0.22 0.57
0.01 0.01 0.01’ 0.02
Values are means of at least 4 livers of different infested animals f S.E. “P < 0.001; bP < 0.005; ‘P < 0.01; dP < 0.02, versus the control value.
f 0.01 f 0.01 ziz 0.02 zt 0.03
224
lism seems to be adapted to a net production of this amino acid needed for tumour growth [lo]. One of the main metabolic fates of methionine is polyamine biosynthesis, so the diminished hepatic contents of this amino acid may partially reflect its preferential shift toward tumour polyamines, paralleling the behaviour of ornithine. Liver cysteine increased in tumour bearing mice along tumour development; the same pattern was observed in plasma [ 111. The results presented here confirm that after tumour transplantation the nitrogen metabolism of the host liver is altered reflecting changes in the free amino acid pools along the tumour development. Acknowledgement This study was supported PB/88/0455 from the Direcci6n InvestigacSn Cientifica y Tknica.
9
10
11
12
13
14
by Grant General de
713-719. Carrascosa, J.M., Martinez, P. and N6tiez de Castro, I. (1984) Nitrogen movement between host and tumor in mice inoculated with Ehrlich ascitic tumor cells. Cancer Res., 44, 3831-3835. Felig, P.. Pozefsky, T., Marliss, E. and Cahill, (1970) Alanine: key role in gluconeogenesis. 167, 1003-4.
G.F., Jr. Science,
Janne, J., Poso, H. and Raina. A. (1978) Polyamines in rapid growth and cancer. Biochim. Biophys. Acta, 473, 241-293. Kawamura. I., Moldawer, L.L., Keenan, R.A., Batist. G., Bothe, A., Jr., Altered amino growth. Cancer Laze, P. (1981)
Quesada. A.R. and NGfiez de Castro, 1. (1989) Nitrogen metabolism in tumor bearing mice. Arch. Biochem. Biophys., 268, 667-675. Olavarria. J.S., Chico, E., GimBnez-Gallego, G. and NdRez de Castro, I. (1981) Effect of ammonium ions on the aerobic glycolysis in Ehrlich ascites tumor cells. Biochimie. 63. 469-475. Pain, V.M. and Garlick. P.J. (1980) The effect of an Ehrlich ascites tumour on the rate of protein synthesis in muscle and liver of the host. Biochem. Sot. Trans., 8,354. Quesada, A.R.. Medina. M.A., Mbrquez, J.. SbnchezJimenez. F. and Ndtiez de Castro, 1. (1988) Contribution
15
by host tissues to circulating glutamine in mice inoculated with Ehrlich ascites tumor ceils. Cancer Res.. 48. 1551-1553. Shapot, V.S. (1980) Manifestations and mechanisms of
16
systemic effect of tumours on the host. In: Biochemical aspects of tumour growth. pp. 124-173. Editor: D.A. Myshne. MIR Publishers, Moscow. Tessitore. L., Bonelli, G. and Baccino. F.M. (1987) Early
References Ardawi. M.S.M. and Newsholme, E.A.(1985) Fuel utilization in colonocytes of the rat. Biochem. J.. 231.
M6rquez, F.J., Quesada, A.R.. Sbnchez-JimPnez, F. and N6fiez de Castro, I. (1986) Determination of 27 Dansyl amino acid derivatives in biological fluids by reversedphase high performance liquid chromatography. J. Chromatogr., 380. 275-283. MBrquez, J., M&es. J .M., Quesada, A.R., Medina, M .A., N(liiez de Castro, I. and SBnchez-JimQnez, F. (1989) Altered ornithine metabolism in tumor-bearing mice. Life Sci., 45, 1877-1884. MBrquez. J., S6nchez-JimLnez, F., Medina. M.A.,
17
18
19
development of protein metabolic perturbations in the liver and skeletal muscle of tumour-bearing rats. Biochem. J.. 241, 153-159. Urdiales. J.L., Medina, M.A., Ndtiez de Castro, I. and SBnchez-Jimhnez, F. (1989) Early systemic effects on the hepatic mitochondria of tumour bearing mice. Cancer Lett.. 44, 179-183. Vincent, M.D. (1985) The clinical problem. In: The molecular basis of cancer, pp. l-35. Editors: P.B. Farmer and J.M. Walker. Croom Helm, London. Wagle, S.R., Morris. H.P. and Weber, G. (1963) Comparative biochemistry of hepatomas. V. Cancer Res., 23. 1003- 1007. Waterhouse, C., Jeanpretre, N. and Keilson, J. (1979) Gluconeogenesis from alanine in patients with progressive malignant disease. Cancer Res.. 39. 1968-1972.
Bistrian, B.R. and Blackburn. G.L. (1982) acid kinetics in rats with progressive tumor Res., 42, 824-829. Amino acids and glucose utilization by dif-
21
ferent metabolic pathways in ascites tumour cells. Eur. J. Biochem., 117, 1925. Levin. L., Gevers. W., Jardine, L., De Guel, F.J.M. and Duncan, E.J. (1983) Serum amino acids in weight-losing
West, J.T. book W.R.
22
patients with cancer and tuberculosis. Eur. J. Cancer Clin. Oncol., 19, 711-715. MBrquez, F.J. (1987) Estudio din6mico de amino6cidos libres en el sistema huesped-tumor en ratones inoculados
Williamson, D.H.. Lopes-Vieira. 0. and Walker, B. (1967) Concentrations of free glucogenic amino acids in livers of rats subjected to various detabolic stresses. Biochem. J.. 104. 497-502.
23
Windmueller. H.G. and Spaeth. metabolism of plasma glutamine Biol. Chem., 249. 5070-5079.
con el carcinoma sity of Mhlaga.
ascitico de Ehrlich. Ph.D. Thesis, Univer-
20
ES., Todd, W.R., Mason, H.S. and Van Bruggen, (1966) General composition of the body. In: Textof Biochemistry, pp. 416. Editors: E.S. West and Todd. The Macmillan Company, New York.
A.E. (1974) Uptake and by the small intestine. J.