ICANCER
RESEARCH
53,5143-5147.
November
i, 1993)
Purine Nucleotide Levels in Host Tissues of Ehrlich Ascites Tumor-bearing Mice in
Different Growth Phases of the Tumor
Werner G. Siems1, Tilman Grüne,Heike Schmidt, Yuri V. Tikhonov, and Alexej M. Pimenov
Herzog-Julius Hospital, Kurhausstrasse 13-17, D-38655 Bad Harzburx [W. G. S.], Clinics of Physiotherapy, Medical Faculty (Charité),Humboldt University, Berlin, D-10098
[T. G., H. S.¡,Germany, and Department of Biochemistry, Central Research Laboratory, N. I. Pirogov 2nd Medical Institute, Moscow, 117437, Russia [Y. V. T., A. M. P.]
so-called normal tissues of the host and in the blood plasma? There
were published data on nucleotide levels and activities of nucleotidemetabolizing enzymes in selected host tissues (liver, lymphocytes,
erythrocytes) of tumor-bearing subjects (14-21). The host-tumor
ABSTRACT
The pool sizes of purine nucleotides, nucleosides, and nucleobases were
measured in the host tissues liver, skeletal muscle, and blood of Ehrlich
ascites tumor-bearing mice during the different periods of tumor growth.
There were large differences of tissue concentrations of these metabolites
between control animals, animals in the logarithmic growth period of the
Ehrlich ascites tumor, and animals in the resting phase of tumor growth.
The ATP concentrations in liver, muscle, and erythrocytes were higher
during the proliferating phase of the tumor compared with the ATP levels
of these organs in healthy animals. In liver and skeletal muscle the ATP
concentration decreased during the transition from proliferating into rest
ing phase of tumor growth. The concentrations of nucleosides and nucleo
bases within the RBC and blood plasma decreased during the logarithmic
growth phase but restored during the plateau period. As well as in the
organs/cells investigated and in the body fluids (plasma, ascites fluid) a
tremendous increase of adenosine concentration during the resting phase
of tumor growth was observed. From changes of total purine ribonucleotide pattern an activation period in the nucleotide metabolic pathways of
liver and skeletal muscle during the proliferating phase of tumor growth
is postulated.
interrelationships in the purine nucleotide metabolism are not well
understood. One can assume that both the liver and the skeletal
muscles are the organs with the highest production of nucleotide
precursors for the whole organism (22). On the other side the RBC and
the body fluids, the blood plasma and in ascites tumor-bearing animals
also the cell-free ascites fluid, are transport media between purincproducing organs and the growing purine-consuming ascites cells.
MATERIALS
AND METHODS
Materials. All reference standards were purchased from Bochringer,
Mannheim, Germany. NH4HiPO4 and KH2PO4 were obtained from Fisher
Scientific Co., Fairlawn, NJ. Acetonitrile and melhanol were purchased from
Merck, Darmstadt, Germany. Tetrahutylammonium
phosphate was from
Waters Associates. Milford. MA.
Sample Preparation. Female ICR mice, weighing approximately 15-20 g,
were used. The cells obtained from mice 7 to 9 days after inoculation of the
tumor were suspended in saline solution to a concentration of 5 x IO7 cells/ml
INTRODUCTION
During the growth of EATC2 there exists a proliferating phase, in
which the number of cells increases quasi-exponentially, followed by
a resting phase, in which the number of cells stays practically con
stant. The growth retardation is not due to an increased rate of cell
death, but is caused by the prolongation of phases of the cell cycle and
the increased share of cells of the resting fraction (G()), whereas the
percentage of S-phase cells is decreased ( 1). The mechanisms of these
changes have been obscure until now. The effects of several growth
stimulating and inhibiting factors have been discussed (2, 3). During
the transition of EATC from the proliferating into the plateau period
of growth there occur morphological changes, i.e., structural deterio
ration and decreased number of mitochondria (4); metabolic changes,
i.e., decreased DNA biosynthesis (5); RNA synthesis, protein synthe
sis (6); protein degradation (7, 8); and changes of the protein pattern
(9) and concentration of other metabolites. A tremendous loss of
intracellular purine and pyrimidine nucleotides, nucleosides, and
bases was measured (4) during the transition of Ehrlich ascites tumor
from the proliferating into the resting phase of tumor growth. These
changes correlate with a decline of the incorporation rates of adenine,
hypoxanthine, adenosine (4), with a decrease of the ATP turnover
(10-12) and changed metabolic flux rates through pathways of ad
enine metabolism (13). The reasons for these changes of the purine
ribonucleotide metabolism are still unknown.
The question which should to be answered by these investigations
was: are the drastic changes of purine pattern in Ehrlich mouse ascites
cells accompanied by deviations of nucleotide concentrations in the
Received 6/24/93; accepted 8/25/93.
The costs of publication of this article were defrayed in part hy the payment of page
charges. This article must therefore he herehy marked advertisement in accordance wilh
18 U.S.C. Section 1734 solely to indicate Ihis fact.
1To whom requests for reprints should he addressed, at Herzog-Julius Hospital.
Kurhausstr.13-17 D-3H655 Bad Harzburg, Germany.
- The abbreviations used are: EATC, Ehrlich mouse ascites tumor cells: HPLC, high-
and then 0.5 ml was inoculated. On the fifth and 12th days after inoculation,
liver, femur muscle, blood, and cell-containing ascites fluid were taken from
anesthetized animals. The mice were anesthetized with diethyl ether. Liver and
muscle were rapidly frozen in liquid nitrogen. Blood and cell-containing as
cites fluid were centrifuged (900 X g, 2 min). Plasma and cell-free ascites
fluid were rapidly separated from the cells. RBC were resuspended in saline
solution before extraction procedure. The Ehrlich mouse ascites cells were
washed and resuspended for nucleotide determinations. The hematocrit
values were measured for blood and erythrocyte suspensions. The reticulocyte content was counted after staining of RBC with isotonic brilliant cresyl
blue solution. The weights of solid organs were measured frozen. Solid
organs were homogenized in saline solution and after that were extracted.
Extraction Procedure. The samples were deprotcinized with ice-cold 6%
perchloric acid, centrifuged for 10 min at 1200 x g and neutralized with
K,CO, (1.3 M).After filtration the samples were stored at -20°C; 10 or 20 fil
were analyzed by HPLC.
HPLC Instrument. An equipment of Waters Associates was used consist
ing of a M510 pump, a U6K Universal Liquid Chromatograph injector, a 254
nm wavelength detector, a variable wavelength detector (adjusted to 280 nm),
a Model 730 data module, a Model 740 data module and a Model 640 system
controller; 5 firn Nova Pak C]K cartridges (100 mm x 8 mm inside diameter)
with a Z-module compression system were used.
Determination of Nucleotides. The determination of nucleotides was per
formed in cells with an isocratic ion pair reverscd-phasc HPLC system. The
buffer contained 10 ITIMNH4H2PO4, 2 ITIMtetrabulylammonium phosphate, and
20% acetonitrile. The flow rate was 2 ml/min.
Determination of Nucleosides and Nucleobases. Two different modifica
tions of HPLC separations published previously (23) were used: a for the
determination of nucleosides and nucleobases in cells, which contain high
concentrations of nucleotides, an isocratic reversed-phase system was used.
The eluent was 7% methanol and 50 mm KH2PO4 (pH 4.1). The high salt
concentration induces the fast elution of high-concentrated rihonucleotides in
performance liquid chromatography.
a tight frontal peak and, therefore, leads to their Chromatographie separation
from nucleosides and nucleobases. (b) For the determination of nucleosides
and nucleobases in body fluids a buffer containing 8% methanol and 10 rriM
KH:PO4 (pH 5.9) was used. To reduce the separation time, especially of
adenosine, which was eluted as a very late peak, a flow gradient was used. The
5143
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PURINE
NUCLEOTIDES
IN HOST TISSUES
flow conditions were 1.5 ml/min from zero time to 6 min, a gradient from 1.5
ml/min to 3 ml/min within 1 min, and a following fi-min run with 3 ml/min.
Thereafter the flow rate was reduced to 1.5 ml/min.
The peak identification was performed by coelution of biological samples
with standard mixtures. Peak identification was confirmed in additional meth
odological studies by diode array detection but was not documented here in
detail.
Statistical Analysis. Significance analysis was performed by means of the
Wilcoxson-Wilcox test for P < 0.05 or P < 0.01.
OF TUMOR-BEARING
MICE
nmoles / g w.w.
3000
2500
++,§§
2000
RESULTS
1500
Table 1 documents nucleotide concentrations of Ehrlich mouse
ascites cells on the fifth and 12th days with the tremendous decrease
of those concentrations at the resting period of tumor growth in
comparison with the proliferation period. In Fig. 1 the liver concen
trations of the adenine nucleotides ATP, ADP, and AMP during the
EATC growth are shown. The day zero represents physiological val
ues of the mouse liver before inoculation of tumor cells. The 5th day
after transplantation of EATC represents the behavior of liver adenine
1000
500
0
0
5
12
EMAT growth, days
Table l Purine nucleotide pattern of EATC cells at the fifth day after inoculation of
ihe tumor (proliferating phase) and at the 12th day (resting phase of tumor growth}
Fig. 2. Concentrations of ATP,ADP, and AMP in skeletal muscle of Ehrlich ascites
tumor-bearing mice in the course of tumor growth. Symbols and dimensions are as in Fig.
1. n = 6 animals for each day. *, P < 0.05 for 5th day values in comparison with zero
Values for nuclcotidc concentrations in nmol/g wet weight, mean ±SD; 14 indepen
dent experiments in each tumor growth period (*, P < 0.05: **, P < 0.01).
values; +, P < 0.05 and + + ,/"< 0.01 for 12th day values in comparison with 5th day
values; §§,
P < 0.01 for 12th day values in comparison with zero values. EMAT,Ehrlich
Proliferating phase
Resting phase
mouse ascites tumor; w.w.,wet weight.
ATPADPAMPGTPGDPUTPUDPATP:ADPATP:AMPGTP:GDPUTPiUDP3047629227799244180704.8413.423.272.57176602275122990.280.780.310.411136±
151310
37150
±
±25291
nucleotide concentrations during the logarithmic phase (proliferating
4770
±
920±212
±
phase) of tumor growth, and the 12th day represents values during the
±23.66
0.497.57
±
1.014.16
±
0.231.67
±
±0.17.1*
nmoles / g w.w.
2000
1500
stationary phase (plateau phase or resting phase) of tumor growth.
One can see that liver adenine nucleotide levels increased during the
logarithmic phase of tumor growth. The ATP concentration increased
by about 20%, the ADP concentration by about 25%, and the AMP
concentration by about 20% as compared with the initial (physiologi
cal) values. On the 12th day after tumor transplantation the ATP, ADP,
and AMP levels of the host liver were again within the range of
physiological values found in the control group. The ATP levels of the
skeletal muscles (Fig. 2) and of the circulating RBC (Fig. 3) increased
during the logarithmic phase of the EATC growth in comparison with
control values, also. Fig. 2 demonstrates the drastic decrease of
muscle ATP concentration during resting phase of tumor growth lead
ing to a value of about 60% of the control value. The values "sum of
adenine nucleotides" reflect in an analogous manner an "activation"
period of host organs of the tumor-bearing animals during the prolif
eration period of EATC growth (Fig. 4). This activation period mea
1000
sured at the 5th day after inoculation of the tumor includes both the
increase of the ATP level and that of ADP and AMP levels. Therefore,
the ATP:ADP and the ATP:AMP ratios are not increased at the 5th day
in comparison with the control values (Figs. 5 and 6). However, there
500
was measured a drastic decrease of these ratios in muscle and erythrocytes at the 12th day, i.e., during the resting phase of tumor growth,
reflecting the unfavorable energetic state of the tissues during the late
period of tumor development.
0
5
12
The data on the guanine ribonucleotide concentrations and the
GTP:GDP ratio are shown in Table 2. The growth phase-dependent
EMAT growth, days
changes of GTP, GDP, GMP(+IMP), and sum of guanine nucleotide
Fig. 1. Concentrations of ATP.ADP. and AMP in the liver of Ehrlich ascites tumorbearing mice in Ihe course of tumor growth. The symbols are: ( * ) ATPconcentration: (•) concentrations in host tissues are much lower in comparison with the
ADP concentration: and (A) AMP concentration. The values are given as nmol/g (w.w.): changes of adenine nucleotide concentrations. The elevation of pool
mean ±SD. The 5th day after inoculation of the tumor represents the proliferating phase
sizes during the proliferating period of EATC growth as seen for the
and the 12thday represents the resting phase of tumor growth. Time zero represents values
ATP does not occur in these metabolites, except a slight tendency for
of animals without tumor cell inoculation (physiological or control value). Number of
experiments. 6 animals in each group. Each single value from one animal was calculated
such behavior in the muscle GTP concentration.
from at least two measurements. Statistical analysis: *, P < 0.05. and **, P < 0.01 for 5th
Figs. 7 and 8 document the changes of nucleoside/nucleobase levels
day values in comparison with zero values; +, P < 0.05 for 12thday values in comparison
with 5th day values. EMAT, Ehrlich mouse ascites tumor.
in the blood plasma and in the erythrocytes. Additionally, Table 3
5144
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PURINE
NUCl.EOTIDES
IN HOST TISSUES
nmoles / g w.w.
OF TUMOR-HI-.ARING
MICE
tumor (see Table 1) are in accordance with observations on other cells.
In hcpatocytes and hepatoma cells, a decline of DNA synthesis (2628), and an increase of DNA catabolism (29), a decrease of purine and
pyrimidine de novo syntheses, as well as enhanced purine and pyrimidine catabolism (15) in connection with a decrease of growth and
proliferation has been found. Nevertheless, the reasons for the drastic
changes in the nucleotide pattern and metabolism remain unclarified.
We now suggest that the nucleotide metabolism of host organs may
modulate the metabolism of tumor cells, e.g., by the supply of nucleosides and nucleobases during tumor growth. Therefore, the investiga
tion of the nucleotide pattern and metabolism of such host organs like
liver and skeletal muscles, which are of major importance for supply
of the whole organism with purine nucleosides and bases, seems to be
of interest.
Nucleotide Concentrations of Host Organs and Nucleoside/
Nucleobase Concentrations of Blood Plasma and Peritoneal Fluid
during Proliferation and Resting Phase of EATC Growth. There
are very few reports on host organ-tumor interrelationships in purine
2000
1500
1000
500
0
5
12
and pyrimidine metabolism. The concentrations of some purine com
pounds in B- and T-lymphocytes of the mouse during growth of solid
EMAT growth, days
Fig. 3. Concentrations of adcnine nuclcoliüesATP, ADR and AMP of circulating RBC
of Ehrlich ascitcs tumor-hearing mice in the different periods of tumor growth. Symbols
and dimensions as in Fig. l. n = 6 animals for each day of tumor transplantation. *, P <
0.05 for 5th day values in comparison with zero values; 4- -t-,P < 0.01 for 12th day values
in comparison with 5th day values; §§.
P < 0.01 for 12th day values in comparison with
zero values. EMAT, Ehrlich mouse ascites tumor; w.w., wet weight.
nmoles / g w.w.
3500
3000
5
12
EMAT growth, days
Fig. 5. ATP:ADP ratio of liver, skeletal muscle, and RBC during Ehrlich mouse asciles
tumor growth. Symbols and number of experiments as in Fig. 4. *. P < 0.05 for 5th day
values in comparison with zero values; +. P < 0.05 for 12th day values in comparison
with 5th day values, §§,
P < 0.01 for 12th day values in comparison with zero values.
EMAT, Ehrlich mouse ascites tumor.
EMAT growth, days
Fig. 4. Sum of adeninc nucleotidcs in liver, skeletal muscle, and circulating RBC of
mice during Ehrlich ascites tumor growth. Symbols of the columns: Liver (•),skeletal
muscle (W}, crythrocytes (D), n = 6 animals for each day. Mean values ±SD. *, P < 0.05
for 5th day values in comparison with zero values; +, P < 0.05 and + + , P < 0.01 for
12th day values in comparison with 5th day values; §.P < 0.05 for 12th day values in
comparison with zero values. EMAT, Ehrlich mouse ascites tumor; w.w., wet weight.
shows the nucleoside and nucleobase concentrations in liver, skeletal
muscle, and cell-free ascites fluid. In all tissues the adenosine level
decreased at the 5th day in comparison with the control value. At the
12th day the adenosine concentrations in the tissues, in blood plasma,
and cell-free ascites fluid markedly increased.
DISCUSSION
Changes in Nucleotide Concentrations and in Flux Rates of
Energy Metabolism in Ehrlich Ascites Tumor Cells during Tran
sition from one Growth Period into the Other. Functional and
morphological deteriorations of cells are often connected with drastic
alterations in the adenine and guanine ribonucleotide pools (24, 25).
The findings of a loss of intracellular nucleotide concentrations during
the transition from proliferating into resting phase of Ehrlich ascites
EMAT growth, days
Fig. 6. ATP:AMP ratio of liver, skeletal muscle and RBC during Ehrlich mouse ascites
tumor growth. Symbols and number of determinations as in Fig. 4. *, P < 0.05 and **,
P < 0.01 for 5th day values in comparison with zero values; +, P < 0.05 and + +,/*<
0.01 for 12th day values in comparison with 5th day values; §,P < 0.05 and §§,
P < 0.01 for 12th day values in comparison with zero values. EMAT. Ehrlich mouse
ascites tumor.
5145
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PURINE
NUCLEOTIDES
IN HOST TISSUES
Table 2 Concentrations of guanyle ribonuclcotides and the GTP:GDP and CTP:GMP
ratios of host tissues of Ehrlich ascites tumor-bearing mice
Concentration values in nmol/g wet weight: 6 animals; 12 measurements; mean ±SD.
(*, P < 0.05 and **, P < 0.01 for 5th day in comparison with zero values; i,P< 0.05
and tt, P < 0.01 for 12th day in comparison with 5th day values; §,P < 0.05 and
§§,
P < 0.01 for 12th day in comparison with zero values).
LiverGTPControl
muscle143
154(100)5th 497 ±
(80)*12th
day
399 ±35
(63)t,§§GDPControl
day
312 ±35
(100)187
±9(100)Erythrocytes159
±33
(107)140
±16(131)**
170 ±26
(305)§.t23
±85
(98)tt31
±33
(100)5th
93 ±23
(91)12th
day
25 ±11
(84)GMPControl
day
78 ±10
(100)24
±3
(77)51
±8
±7(165)t,§32
(100)34
±5
(148)45
±6
(196)§8
±5
(100)5th
43 ±15
(81)12th
day
35 ±7
(65)§Sum
day
28 ±6
GN"Control
192(100)5th 633 ±
(82)12th
day
519 ±53
(66)§GTP:GDPControl
day
418 ±51
13(100)32
±
(100)22
±11
(69)206
±5
(100)15
±1
±5(188)*40
(500)t,§§190
±9
(100)243
±25
(118)*213
±35
±45(103)t4.61
39(100)219
±
(115)570
±37
(300)§§6.91
±99
1.665th
5.34 ±
0.4112th
day
4.69 ±
0.45§GTP:GMPControl
day
4.00 ±
0.297.79
±
0.67**2.75
±
0.65tt,§§4.47
±
1.435.00
±
0.7610.78
±
2.89t,§19.88
±
OF TUMOR-BEARING
MICE
In liver and skeletal muscles of Ehrlich ascites tumor-bearing mice
a significant increase of adenine nucleotide concentrations was ob
served at the fifth day after inoculation of the tumor, i.e., during the
proliferating phase of tumor growth. The ATP, ADP, and AMP con
centrations were increased by about 20 to 25%. We called this phe
nomenon activation period in adenine nucleotide metabolism. In this
period of its growth the tumor consumes an enormous amount of
nucleotide precursors for cell proliferation, and DNA and RNA syn
theses. The high demand of nucleosides and nucleobases for such
syntheses in the tumor cells leads to the decrease of purine nucleoside
and nucleobase concentrations in the blood plasma (see Fig. 8) and in
the peritoneal fluid surrounding the ascites tumor cells, also. Even
under conditions of increased flux rates in salvage and de novo syn
thesis pathways in liver and muscles the increased nucleoside and
nucleobase supply by these organs cannot prevent the decrease of
plasma levels of purine nucleosides and nucleobases. The observation
of an activation of purine nucleotide metabolism in liver and skeletal
muscles is in accordance with results of Weber et al. (14, 15), who
described an increase of activities of enzymes of the de novo syntheses
nmoles / ml
0.285.84
±
4.1311.33
±
3.585th
11.56 ±
0.50*6.36
±
1.7312.13
±
1.0012th
day
11.40 ±
±2.13nmoles
1.50§concentrations.485
±
1.25" day
11.14 ±
Sum GN, sum of guanine nucleotideSkeletal
w.w.8642A-k+
/g
3
,§§J-*is!+
0
2)~Vi>
5
12
EMAT growth, days
0
5
Fig. 8. Concentrations of purine nucleosides and nucleobases in the blood plasma. For
symbols see Fig. 7. *, P < 0.05 for 5th day values in comparison with zero values; + + ,
P < 0.01 for 12th day values in comparison with 5th day values; §,P < 0.05 and §§,
P
< 0.01 for 12th day values in comparison with zero values. 1) + +, §for O; ++, §§
for
*. EMAT. Ehrlich mouse ascites tumor.
12
EMAT growth, days
Fig. 7. Concentrations of nucleosides and nucleobases in circulating erythrocytes. As
symbols were used: (*) adenosine, (O) inosinc, (•)adenine, and (A) guanosine. Six
animals for each value: mean ±SD. *, P < 0.05 for 5th day values in comparison with
zero values, + + , P < 0.01 for 12th day values in comparison with 5th day values;
§,P < 0.05 and §§,
P < 0.01 for 12th day values in comparison with zero values.
1) ** for O; ** for •¿;
2) +4- for O; §for •¿
EMAT,Ehrlich mouse ascites tumor; w.w.,
wet weight.
Table 3 Nucleoside and nucleobase concentrations in liver, skeletal muscle, and
cell-free ascile* fluid
Values in nmol/g wet weight or nmol/ml ascites fluid, respectively; 6 animals, 12
measurements; mean ±SD. Symbols for statistical analysis as in Table 2.
hepatomas has been described (30). These authors found a tremendous
decrease of xanthine and hypoxanthine concentrations in the B- and
T-lymphocytes during the phase of exponential growth of the tumor
(5th to 8th day after inoculation of the tumor) due to a reduction of the
adenosine deaminase activity of those cells. Furthermore, under such
conditions also an accelerated salvage pathway should contribute to
the decreased concentration of hypoxanthine.
A preliminary study of the purine pattern of circulating erythrocytes
in mice bearing the Ehrlich ascites tumor was published previously by
this laboratory (20, 21).
muscle2.6
fluid0.01
±0.11.7
±0.3*5.1
day12th
dayAdenineControl5thl.ltt,§§0.9
±
0.72.5
±
0.75.3
±
1.7tt,§§1.3
±
0.000.03
±
O.OOtt1.2
±
0.61.5
±
day12lh
±0.30.7±0.1t4.7
dayInosineControl5th
±0.11.6
±0.73.3
1.8t,§§2.4
±
0.67.2
±
±1.1*5.4
day12th
dayGuanosineControl5th
Of6.9
±1.
±1.02.5
1.14.0±1.8t,§4.5
+
AdenosineControl5th
1.15.6
±
1.15.2
±
0.93.6
±
±2.22.0
day12th
±0.6t,§Ascites
±0.6f,§Skeletal
dayLiver1.9
0.31.3
+
0.21.2
+
0.11.3
+
0.10.8
+
0.10.5
+
±0.11
5146
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PURINE
NUCLEOTIDES
IN HOST TISSUES
of purines and pyrimidines during the proliferation phase of tumor
growth. Furthermore, activation of the salvage pathways occurred (14,
15). That may explain our results on the increase of adenine nucleotide
concentrations in liver and skeletal muscle. The activation in adenine
nucleotide metabolism occurs in the same period of the tumor growth
in which further pathways are also stimulated, at least in the liver, like
protein syntheses (31, 32), as discussed above. Concerning the liver
one may speak on a general functional activation period with the aim
of increased substrate supply (for ATP production) and precursor
supply (for syntheses) in the early phase of tumor growth. Besides
increased glucose supply, increased utilization of lactic acid (see pH
homeostasis), etc., the liver obviously is responsible for supplying
tumor cells with precursors for nucleotide metabolism and nucleic
acid syntheses during logarithmic tumor growth, also.
The decrease of purine nucleotide concentrations of liver and
muscle at the 12th day after transplantation of the tumor in compari
son with the 5th day can be discussed with regard to changes during
the cancer cachexia at the resting phase of the tumor growth.
In contrast to the adenine nucleotides reflecting an activation period
in the nucleotide metabolism in liver and muscle, the concentrations
of the guanine nucleotide decrease during the whole course of the
Ehrlich ascites tumor growth. The change of the concentration of a
metabolite is always determined by the difference between influx
rate(s) and outflux rate(s) in the pool. Therefore, the consumption of
guanine nucleotides, nucleosides, and nucleobases must be higher
than their generation in the host organs (de novo synthesis plus sal
vage pathways). In the case of the adenine nucleotides, nucleosides,
and bases this imbalance obviously can be prevented. The consump
tion of guanine nucleotides by tumor cells is about approximately
equal with the consumption of adenine nucleotides because of the
equal number of adenine and guanine bases in DNA and RNA. Ob
viously, the increase of guanine nucleotide consumption is much
higher than the capacities for guanylate biosynthesis limited by the
activity of the NAD-dependent IMP dehydrogenase and the salvage
pathway for GTP formation. Nevertheless, also in the pathways of the
guanylate metabolites, the generating flux rates in the host organs liver
and muscle are suggested to be increased during the proliferating
period of EATC growth. But that could only be demonstrated by tracer
kinetic experiments.
The report on the i.p. injection of AMP and ATP in tumor-bearing
animals preventing drastic decrease of the body mass during tumor
growth (33) can be interpreted in the sense that by degradation of ATP
and AMP adenosine and adenine are formed, leading to a nutritive
unloading of the substrate supplying host tissues under these condi
tions.
Finally, it is suggested that new insights into the variations of
nucleotide metabolism of host tissues and of tumor cells in different
growth phases of tumor may open new diagnostic and therapeutic
approaches because of the medical importance of the transition of
tumor cells from one growth period to the other.
ACKNOWLEDGMENTS
Theskillfultechnicalassistanceof ManuelaJakstadtis thankfullyacknowl
edged.
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OF TUMOR-BEARING
MICE
4. Siems. W., Schmidt, H., Werner, A., Uerlings, I., David, H., and Gerber, G. Changes
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Downloaded from cancerres.aacrjournals.org on June 17, 2017. © 1993 American Association for Cancer Research.
Purine Nucleotide Levels in Host Tissue of Ehrlich Ascites
Tumor-bearing Mice in Different Growth Phases of the Tumor
Werner G. Siems, Tilman Grune, Heike Schmidt, et al.
Cancer Res 1993;53:5143-5147.
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