Download Lactate and Pyruvate Metabolism and Reducing

(CANCER
RESEARCH
34, 872-877, April 1974]
Lactate and Pyruvate Metabolism and Reducing Equivalent
Transfer in Ehrlich Ascites Tumor'
Joseph Katz, Karl Brand,2 Sybil Golden, and David Rubinstein
Cedars-Sinai Medical Research Institute, Los A ngeles, California 90029
SUMMARY
teristics
(9, 10). These strains
reportedly
differ greatly in
GPDH3 activity (5, 9—1
1, 17, 18). It appeared to us that a
The metabolism of labeled lactate and pyruvate by three
strains of Ehrlich ascites tumor cells, H, HL, and W, was
studied. The major products from the acids were CO2,
amino acids (mainly alanine), and acetate. HL-cells are
characterized
by a high'4C yield in amino acids, and
W-cells are characterized by their large acetate production.
The rate of lactate and pyruvate oxidation was equal in Hand HL-cells, but W-cells oxidized pyruvate faster than
lactate. Phenazine methosulfate stimulated lactate oxida
tion in W-cells but not in the other two strains. Aminooxy
acetate (2 mM) depressed ‘4C
yields in amino acids by 80 to
90%. It had no effect on lactate oxidation in H- and HL
cells, but depressed it in the W-cells by about 40%. The
results suggest that the oxidation of cytosolic reduced
nicotinamide adenine dinucleotide is rate limiting in the
oxidation of lactate by W-cells but not the H- and HL-cells.
INTRODUCTION
Ascites cells, in common with many tumors, have a fairly
high endogenous rate of oxygen uptake, and they oxidize
lactate and pyruvate at substantial rates (20). The metabo
lism of lactate differs from that of pynuvate by the produc
tion of cytosolic NADH, which must be neoxidized either
in the cytosol or by hydrogen transfer to mitochondnia.
Comparative studies with the 2 acids have been of value to
study the role of reducing equivalent transport in several
organs. Several schemes for the oxidation of cytosolic
NADH have been proposed, the most prominent ones being
the aspartate : oxaloacetate : malate cycle (3) and the a-glyc
erophosphate
shuttle (4), but the mechanism of reducing
equivalent transfer between cytosol and mitochondnia is
not yet established.
In 1905, Ehnlich isolated a line of mouse ascites tumor
cells that have been propagated ever since in many labora
tories. The cells have differentiated
into several strains,
distinct in chromosomal, growth, and biochemical charac
‘Supported
by USPHS
2 Present
address:
Institute
Grants 5ROIAN
for
Physiological
1260405 and RR
Chemistry,
The
W. Contrary to published reports, we found the GPDH
levels in all 3 strains to be similar. Our results indicate that
oxidation of cytoplasmic NADH is rate limiting in the
W-cells but not in the other strains. We provide some evi
dence that transamination plays some role in reducing
equivalent transfer in the W strain, but in the other strains
the role of transamination
appears to be minor.
MATERIALS AND METHODS
Ascites Cells. Hyperdiploid
H-cells and hypodiploid
Lettrémutant HL-cells were obtained from Dr. T. Terra
nova, Universita Catholica, Rome, Italy. A hypotetraploid
strain, W, was obtained from the Institute of Experimental
Pathology of the Bayer AG. Werke in Wupperthal-Elber
feld, Germany. According to Dionisi et al. (5), the activity
of GPDH in the H strain was 25 times that in the HL strain.
According to Letnansky (9, 10) and Letnansky and KIc
(I I), who studied it extensively, the W strain had virtually
no GPDH activity.
The cells were propagated by i.p. injection into mice and
were harvested 7 to 10 days after inoculation.
Incubation. The peritoneal fluid was centrifuged and the
cells were washed 4 to 5 times in 20 volumes of Krebs
Henseleit bicarbonate buffer (19). Packed cells, 0.2 to 0.4
ml, were incubated in 2 ml of this buffer, in an atmosphere
of 95% 02-5% CO2 in 50-mi Erlenmeyer flasks, closed
with rubber serum caps, with center wells containing a vial
for CO2 collection. The incubation at 38°was terminated
by injection through the rubber cap of 2.0 ml phenethyla
mine : water (1 : 1 v/v) into the vial and 0.5 ml 30% HC14
into the medium.
Shaking
was continued
for 1 hr for ‘4C02
collection.
Fractionation. The methods were essentially those used
with other tissues (8). The medium and cells were trans
05468.
Univer
sity Erlangen-Nurnberg, 8520 Erlangen, Egerlandstrasse 7, Germany.
Received November 5, 1973; accepted January 3, 1974.
872
comparison of the metabolism of lactate and pyruvate in
such strains would be of interest. We describe the effects
of PMS and the transaminase inhibitor AOA on the me
tabolism of these acids by 3 strains, namely, H, HL, and
3 The
abbreviations
used
are:
GPDH,
a-glycerophosphate
dehydrogen
ase; PMS, phenazine methosulfate; AOA, aminooxyacetic acid.
CANCER RESEARCH
VOL. 34
Downloaded from cancerres.aacrjournals.org on April 30, 2017. © 1974 American Association for Cancer Research.
Reducing
ferred quantitatively into tared l0-ml graduated centrifuge
tubes and made to volume, and the supernatant fluid was
neutralized with potassium bicarbonate. The cell residue
was washed and dried to obtain the dry weight of the cells.
Aliquots of the neutralized extract were used for assay of
lactate, pyruvate,
and acetate; about one-half passed
through tandem columns (I x 8 cm) of Amberlite CG 120
(100 to 200 mesh) in H@ form, followed by a 1- x 10- cm
column of Dowex AG 1-8 (100 to 200 mesh), in the acetate
form. The columns were eluted with water and a 25-ml
fraction was collected. This fraction consists of neutral
compounds. The columns were separated, and the cation
column was eluted with 20 ml of 2 N NH4OH. The am
monia extract contains the amino acids fraction. The anion
column was eluted with 100 ml 0.25 N acetic acid, which
eluted acetate and most of the /@-hydnoxybutyrate, followed
with 50 ml 1 N formic acid to elute lactate, followed with
70 ml of 4 N formic acid to elute pyruvate. The 4 N for
mate fraction contained some phosphate esters. Finally, the
column was eluted with 25 ml of I N ammonium fonmate.
This removes most of the phosphate esters and di- and tn
carboxylic acids. The cell residue was dissolved with warm
ing in 0. 1 N KOH and the extract was made to volume. The
14Cin this fraction represents the insoluble residue.
Enzyme Assays. Washed cells were stored frozen. The
cells were thawed, suspended in 10 volumes of buffer, and
treated
sonically
at maximum
efficiency
(Branson
sonifier)
4 times for 30 sec with intermittent cooling. The super
natant was used for enzyme assay according to the methods
of Shonk and Boxer (15) and of Bergmeyen (2).
Isotopes. L-Lactate and pyruvate, uniformly labeled
with ‘4C,were purchased from New England Nuclear
(Boston, Mass.) or Amersham/Searle
Corp. (Chicago,
Ill.). The lactate was isotopically pure, but pynuvate con
tamed variable amounts ofp-pyruvate.
When available, the
pyruvate was purchased in 5 jzCi vials, and all of it was
used immediately upon opening, avoiding storage. Alter.
natively, the labeled pyruvate was taken up in 0. 1 N HC1,
divided into lots that were stored at —20°,and was used
immediately upon thawing. Nearly all commercial batches
of labeled pyruvate contained some p-pyruvate, which in
creased upon storage. The labeled pynuvate was analyzed
by column chromatography with every experiment, and the
activity in the pyruvate fraction was determined. The utili
zation of ‘4Cwas calculated from the activity of this frac
tion. The p-pyruvate seems to be metabolically inert. Solu
tions of unlabeled pyruvate were prepared freshly before
use.
Other Assays. Acetate was assayed according to the
method of Shulman and Wood (16). We gratefully ac
knowledge the gift of acetokinase and succinyl-CoA from
Dr. Marvin Shulman of Western Reserve University. Other
assays were transformed
enzymatically
according to the
method of Bergmeyer (2). Pyruvate assays were performed
promptly after neutralization of the extract. The assay of
‘4C
fractions by liquid scintillation was as described else
where (8).
Results were expressed as @tmoles or @iatoms carbon
per 100 mg, dry weight. One ml of packed H- and HL-cells
Equivalents
Transftr
in Ascites
Tumor
corresponds to 100 to I 10 mg, and I ml of W-cells come
sponds to 120 to 130 mg, dry weight.
RESULTS
Lactate and Pyruvate. In Table I, the metabolism of the
2 acids by the 3 strains, H, HL, and W, is compared. The
HL-cells are characterized by their high ‘4Cincorporation
into amino acids, but otherwise the pattern in HL- and
H-cells is similar. Very little pyruvate accumulates in the
W-cells. The point of greatest interest is that in H- and
HL-cells the utilization of lactate is nearly equal to that of
pyruvate, whereas in the W-cells incorporation of pynuvate
into products was I .5 to 2 times that of lactate.
Further information
on the pathway of the pyruvate
moiety in HL- and W-cells is provided by the use of spe
cifically labeled acids (Table 2). Labeled
formed, as expected, solely from carbon atoms
amino acids from pyruvate in both strains
equally labeled from all 3 carbons, indicating
acetate was
2 and 3. The
were nearly
the predomi
nant direct formation of the amino acids from pyruvate.
Alanine was indeed the predominant labeled amino acid.
However, with lactate in W-cells, the incorporation of the
canboxyl carbon into amino acids was much less than that
of C-2 and C-3, indicating that in this case the amino acids
are derived mainly by transamination
of keto acids formed
in the Knebs cycle. The yield of amino acids in the W-cells
is small and the pyruvate concentration is low, so that it is
likely that the pyruvate formed from lactate is preferen
tially oxidized. This conclusion is supported by the finding
of higher ‘4C02
yields from C-3 over C-2 of the lactate.
This would occur only if there is some outflow from the
Knebs cycle.
Of special interest is the formation of acetate. Its identity
was confirmed by Ducleaux distillation (I). Acetate is
rarely if ever the metabolic end product in mammalian
tissues, but Hepp et al. (7) observed its formation in several
types of tumor cells. The highest yield among the tumors
studies was in the tetraploid Ehnlich strain. The W strain is
also tetnaploid. Calculations indicate that in these cells the
acetyl-CoA is predominantly hydrolyzed, and only a small
fraction is oxidized in the Knebs cycle. The amount of ace
tate, as determined by analysis, exceeded that calculated
from the “Cyield, especially in the W-cells, probably due
to acetate formation from endogenous lipid.
Chart I illustrates the kinetics of lactate and pyruvate
utilization by the HL- and W-cells. Amino acid production
levels off at about 30 to 50 mm. It is likely that ‘4Cincor
ponation into amino acids is by transamination
between
pyruvate and endogenous amino acids, and the reaction is
an exchange, rather than net synthesis, of amino acids. In
the HL-cells, acetate formation
decreases
with time,
whereas in the W-cells acetate formation from pyruvate is
much higher and linear until the substrate is nearly ex
hausted.
Effect of PMS. The more rapid utilization of pyruvate
than of lactate by W-cells suggested that oxidation of cy
tosolic NADH was rate limiting. If this is the case, the
APRIL 1974
Downloaded from cancerres.aacrjournals.org on April 30, 2017. © 1974 American Association for Cancer Research.
873
Joseph Katz, Karl Brand, Sybil Golden, and David Rubinstein
Table I
Metabolism of lactate and pyruvate by 3 strains of Ehrlich ascites cells
Thirty-five to 43 mg cells were incubated
in 2 ml bicarbonate buffer for I hr with 10
lactateH
or pyruvate, uniformly labeled with “C.mM
WLactate
HL
Pyruvate
LactatePyruvateUptake:'
Lactate
Pyruvate
mg)Lactate
analytical (pmoks/100
—18'
+1.5
17
Pyruvate
Net
@
Substrate used
Lactateorpyruvateformed
19
1.5
CO2
Acetate
Amino acids
Residue
Others
Total
25
9
15
6
I
56
Lactate or pyruvate formed
Other products
Isotope recovery
3.6
39
99
a
Metabolism oflactate
formation;
+7.5
—23
—25
+2.5
18 32Uptake:21
26
7.0
6.0Recovery
31
II
13
4
1
60
14
44
104
+11
—35
24
—15
+0.5
15+5.5
isotopic (j@moles/100 mg)
24
36
3.0
10
of(jig/atoms
26
6
33
6
71
94%
carbonll00
33
7
30
5
—37
16
0.535
mg) in
18
17
9
4
75
4842
added “C
5.3
18
42
40
101
105
1
25
10211
31
16
4
1
55
103
—, uptake.
Table 2
and pyruvate labeled with “C
in carbon atoms 1, 2, or 3 by strains HL and W ofEhrlich
asciles tumor cells
Twenty-five to 40 mg of cells were incubated in 2 ml of buffer, 14 to 20 mM in lactate and pyruvate, specifically labeled; incubation period, 2 hr.
Experiments with pyruvate and lactate were made with different cell preparations.
or formation (jimoles/l00 mg)Isotopic,
(%)LactatePyruvateNetAcetateCO,AcetateAmino
specific yield
acidsResidueHLPyruvateU-'4C3719368Pyruvatel-'4C+10—24148.0660.5304Pyruvate2-'4C3325347LactateU-―C4973212Lactate
StrainSubstrateLabelaAnalytical,uptake
8
.62
+234
729
Lactatel-―C 2-'4C318
480.5
358
10Lactate3-―C4083715WPyruvateU-'4C4930165Pyruvatel-―C+10—605023770.5176Pyruvate2-'4C2547197LactateU-―C5728510Lactate
Lactatel-―C 2-'4C—31+
12Lactate3-―C3642616
a
@,uniformly
.312296
361
43
labeled.
addition of an auto-oxidizable
hydrogen acceptor such as
PMS should stimulate the utilization of lactate but not of
pyruvate. Indeed, PMS had little effect on pyruvate me
tabolism in concentrations
up to 50 @M,when it became
inhibitory. The effect on lactate is illustrated in Table 3. In
H- and HL-cells, PMS had no effect on CO2 production,
874
480
and there was a small, barely significant stimulation of ace
tate production. In the W-cells, on the other hand, the
oxidation of lactate was nearly doubled. It appears that
only in the W-cells is the oxidation of cytoplasmic NADH
rate limiting.
The Activity of GPDH. According to Dionisi et al. (5),
CANCER RESEARCH VOL. 34
Downloaded from cancerres.aacrjournals.org on April 30, 2017. © 1974 American Association for Cancer Research.
Reducing
E
II)
0
60
HL
0 CO2
S AMINO
E
8
Tumor
U AMINO
6 ACETATE
40
tate but not of pyruvate. In Table 5, the effect of AOA on
oCO2
ACIDS
ACIDS
A ACETATE
UVA@
S
01
0
Transfer in Ascites
mitochondria transaminates with glutamate, and the aspan
tate and a-ketoglutarate
pass into the cytosol where, by a
2nd transamination,
oxaloacetate and glutamate are
formed. The oxaloacetate is reduced to malate, and it and
glutamate transferred to the mitochondria where malate is
oxidized to complete the cycle. The system may serve to
transfer reducing equivalents and carbon between cytosol
and mitochondnia. Rognstad and Katz (1 3) introduced the
transaminase
inhibitor AOA to test for the operation of
this type of transfer, and it has been widely used (12, 14,
21). If transamination
plays a role in the oxidation of cyto
plasmic NADH, AOA should inhibit the oxidation of lac
8
@
Equivalents
20
@@:—.-----@
•
@—.
0
0
30
60
90
120
0
30
60
MINUTES
90
120
MINUTES
Chart I. Kinetics of lactate and pyruvate utilization by HL- and W
cells. Upper half, pyruvate used as substrate; lower half, lactate used
as substrate. Twenty-three mg of HL- and 29 mg of W-cells (dry weight)
were incubated in about 2 ml of incarbonate buffer (see text). Concentra
tion of lactate and pyruvate was 16 mM; 22 and 38%, respectively, was
utilized by the HL-cells, and 30 and 61%, respectively, was utilized by
the W-cells.
Table 3
Efftc: of PMSlactateand
on
2 and acetate formation from glucose,
“CConcentrationpyruvate uniformly labeled with
zsi;incubation,
of lactate and pyruvate, 10 mM; of PMS,
I hr (same experiments as in Table 2).Pyruvate
atoms)Lactate
20
(@zg
carbon/l00
mg)Strain
the metabolism of the acids by the HL- and W-cells is illus
trated. The transaminase
inhibition was manifest in the
sharp drop in the ‘4Cyields in amino acids. Utilization of
pyruvate was not much changed, because the increase in
CO2 and acetate largely compensated for the depression
in ‘4C
incorporation into amino acids and residue (mainly
proteins). In HL-cells in the presence of AOA, the forma
tion of CO2 and acetate was not greatly altered or in
creased, but with W-cells there was a significant depression
in the CO2 yield and a marked depression in acetate. Re
suits with H-cells resembledthosewith HL-cells. It appears
that transaminase
has, if any, a minor role in cytosolic
NADH oxidation in H- and HL-cells, but its role in the W
cells is of greaten importance.
Metabolism of Glycerol and Fructose. Glycerol is metab
olized at a significant rate by HL-cells, but its utilization by
W-cells is one-tenth that of the HL strain (Table 6). This
may reflect differences in the level of glycerol kinase or in
glycerophosphate
dehydrogenase.
The kinase level in the
W-cells was about one-half ofthat in the HL-cells (Table 5).
Letnansky (10) reported that W-cells could not utilize
fructose at all, and this strain appeared unique in this ne
spect.We howeverfound the uptake ofglucose and fructose
to be very similar.
SumH
PMS
CO2
Acetate
Sum
CO2
Acetate
25
25
9
14
34
39
31
29
II
13
40+
—
26
26
6
9
32
35
33
33
7
10
73+
—
18
28
17
37
35
62
42
40
31
39
42+
—
42HL
43W
Table 4
Enzyme assays on 3 strains of ascites cells
For the assay from Vienna, see Ref. 9; for that from Rome, see Ref. 4.
The Los Angeles data are an average of determinations of 2 batches of
rats,
79
GPDH activity in H-cells was 25 times that in the HL-cells.
According to Letnansky (9, 10) and Letnansky and KIc
(1 1), the GPDH content of W-cells was negligible. How
ever, our assay of this enzyme did not agree with these
reports. As shown in Table 4, we found no great differences
which
by from
10 to other.@imoles/hr/mg
20% from each
enzymesStrain
GPDHH
protein with
Laboratory
LDH'
HK
14HI
Vienna
Rome
Dortmund
Los Angeles
250
301
160
220
12.6
5.6
6.5W
Rome
Dortmund
Los Angeles
200
145
70
10
4.8
Vienna
410
1.4
Dortmund
137
8.3
Los Angeles
147
4.0
in the levels of GPDH in all 3 cell lines. The activity ratio
LDH :GPDH in HL-cells was reported to be about 150 in
Rome, but was about 10 by our assay. This ratio for the
W-cells was 1600 in Vienna, as compared to about 20 in our
assays. The level of the GDPH was, in our assays, of the
same order as that of hexokinase.
The Role of Transamination
in Reducing Equivalent
Transport. In the scheme of Borst (3), oxaloacetate in the
differed
a LDH,
lactic
dehydrogenase;
HK,
GK
2.4
hexokinase;
71
31
15.9
1.6
1.4
15
1.2
0.25
7.2
0.6
GK,
6.0
glycerol
APRIL 1974
Downloaded from cancerres.aacrjournals.org on April 30, 2017. © 1974 American Association for Cancer Research.
kinase.
875
Joseph Katz, Karl Brand, Sybil Golden, and David Rubinstein
Table 5
Effect of A OA on pyruvate and lactate metabolism by HL and
strains of Ehrlich ascizes tumor
HL and W-cells. 46 and 55 mg (dry (dry) cells, respectively,
withUptake were incubated with 2 ml buffer, 15 mr@@i
in lactate or pyruvate,uniformly
‘4C,
for 2 hr. Aminooxyacetate concentration, when present, 2 mM.W
or formation
(@tmoles/I00 mg)
inhibi
tion of
Pyruvatea
Lactate°
Acetateb
—
+
45
43
—28
—27
+4.5
+5.0
8.0
12
28
3313
—
+
55
43
+3.0
+0.8
5.5
3.5
39
5312
Pyruvate
Pyruvate
1185Lactate
—
+
89
81
—47
—43
Lactate
—
+
67
40
+0.5
+0.5
AOA
Pyruvate
Pyruvate
688Lactate
6726W Lactate
@
a By elution
from columns.
1, Determined
Lactate
uptake
—29
—23
+9.5
+7.0
24
20
—31
—18
as determined
ofglycerol
Table 6
by strains of Ehrlich ascites tumors
Thirty to 40 mg of cells were incubated for 2 hr in 2 ml buffer with 9 to
12 msi glycerol uniformly labeled with “C.
14C
incorporation mg)CO2
in (natoms carbon/l00
+TotalStrainLactatePyruvateacetateAmino
acidsutilizedHL4.0.57.06.520W0.70.20.50.51.9
DISCUSSION
The Levels of GPDH and the a-GPD H Cycle. Terranova
et al. (17, 18) and Dionisi et al. (5) have used the H and
HL strain in their studies. They suggested, from studies on
oxygen uptake with or without rotenone, that the a-glyc
erophosphate
cycle plays a role in reducing equivalent
in the H- but not the HL-cells.
They correlated
this
with the GPDH level in 2 strains. We could not detect such
a difference and find the GPDH activities to be of the same
order as hexokinase.
In general, the levels of mitochondrial
a-glycerophos
phate oxidase are less than those of the cytoplasmic de
hydrogenase, and in the operation of the shuttle this oxidase
is likely to be rate limiting. Dionisi et al. (5) found the level
of the oxidase
low
and
quite
similar
in H-
and
HL-cells
(0.25 and 0.48 unit), compared with 3 1 and I .4 units, re
spectively, for the DPNH-linked
enzyme. In view of this
it is difficult to see why oxidation via the glycerophosphate
cycle in these 2 cell lines should have differed in their ex
peniments.
The reasons for the difference between the GPDH levels
found in our study and those of the Rome and Vienna lab
876
53
5542
9.5
2.5
analytically
CO3“C
Acetate
54
4624
in close agreement
17
9.5
51
4.5
with column
acids
oxidationAmino
27
12
6
674
23
2.5
9.0
2.083
20
5.5
3.5
0.5
10
6.5125
9.0
2.090
5242
values; see text.
analytically.
Utilization
transfer
incorporated
(jzg/atoms carbon/lOO mg) inTotal%
% added
‘4C
utilized
Strain
Substrate
ResidueHL
labeled
oratories are puzzling. It would be of considerable interest
if ascites strains with confirmed low levels of glycerophos
phate dehydrogenase could be found.
Reducing Equivalent Transfer. Our results indicate that
the oxidation of cytosolic NADH is rate limiting in the oxi
dation of lactate in the W strain, but not in 2 other strains.
The inhibition by AOA suggests that a transaminase-cata
lyzed pathway plays a significant role in the oxidation of
cytosolic NADH in the W-cells.
Aminooxyacetate
in concentrations
from 0.2 to 1 mM
has been used as a probe for transaminase-catalyzed
carbon
and reducing equivalent transfer across the mitochondnial
membrane in kidney (12, 13), liver (21), and heart (14).
High concentrations
of AOA are required completely to
inhibit transamination,
and the compound may not be en
tirely specific and affect other sites. Great caution is needed
in interpreting such results and, by themselves, they are
not conclusive. However we observed no inhibition of CO2
and acetate production from pyruvate, and a significant
inhibition of oxidation from lactate was observedonly with
lactate in the W-cells. While transamination
may play a
role in the oxidation of cytosolic NADH in the W-cells, it is
not likely
that this is the sole pathway
lent transfer
for reducing
equiva
in this strain.
In insects,the role of the a-glycerophosphat.e
cyclefor
the oxidation of cytosolic NADH seems established (4)
but, at present, there appears to be no convincing evidence
for or against the operation of this scheme in mammalian
cells. Further study is required to provide firm expenimen
tal support for the various schemes proposed for reducing
equivalent transport.
The Formation of Acetate. The accumulation of acetate
is rare in mammalian cells. An interesting aspect of the
metabolism of ascites cells was the high yield of acetate.
This is formed to some extent from glucose, but more so
from
lactate
and pyruvate.
In the tetraploid
strain
CANCER RESEARCH
W, ace
VOL. 34
Downloaded from cancerres.aacrjournals.org on April 30, 2017. © 1974 American Association for Cancer Research.
Reducing
tate is the major product of the metabolism of the acids.
The formation of acetate by tumor tissue was suggested by
Elliot in 1937 (6). This received little attention until the
thorough study of Hepp et a!. (7). They found low levels of
acetokinase and a high level of acetyl-CoA deacylase in a
numberof tumors.In normaltissues,the activityof aceto
kinaseexceededthat of the hydrolase.
Our results indicate considerable
differences
these strains
as mutations
9. Letnansky,
10.
of the original
stock, but there is no informationto relate the metabolic
12.
13.
14.
REFERENCES
15.
1. Barker, H. A. Methods Enzymol. 3: 379-381, 1962.
2. Bergmeyer, H. U. Methods of Enzymatic Analysis. New York: Aca
demic Press, Inc., 1965.
3. Borst, P. Hydrogen Transport and Transport Metabolites. In: P. Karl
son (ed.), Functionelle und Morphologische Organization der Zelle,
pp. 137—158.Heidelberg, Germany: Springer-Verlag, Inc., 1963.
4. Bucher, T. A., and Klingenberg, M. Wegen des Wasserstoffs in der
Lebenden Organization. Angew. Chem., 70: 552-570, 1958.
5. Dionisi, 0., Cittadini, G., Galeotti, T., and Terranova, T. The Role of
the a-Glycerophosphate Shuttle in the Reoxidation of Cytosolic
NADH in Ehrlich Ascites Tumor. Biochim. Biophys. Acta, 216: 7179, 1970.
6. Elliot, K. A. C., and Greig, M. E. Metabolism of Lactate and Pyruvic
Acid in Normal and Tumor Tissue. The Formation of Succinate. Bio
chem. J.,31: 1021—1032,
1937.
7. Hepp, G., Prusse, E., Weiss, H., and Wieland, 0. Essigsaure als End
product des aeroben Krebsstoffwechsels. Biochem. Z., 344: 87-102,
in Ascites
Tumor
1966.
in the met
pattern to chromosomal changes. The discrepancies be
tween the enzyme profiles reported by other laboratories
and our findings may indicate that the “mutated―
strains
reverted back to the original type. Collection and preserva
tion of the various strains should be of interest.
Transftr
8. Katz, J., and Wals, P. A. Pentose Cycle and Reducing Equivalents in
Rat Mammary-Gland Slices. Biochem. J., 128: 879-899, 1972.
abolicpatternof the Ehrlichascitesstrains.Letnansky(9— 11.
11) considered
Equivalents
K. Unterschiede
der
Fermentmuster
verschiedener
Stämmedes Ehrlich-Ascites Tumors. Z. Krebsforsch., 70: 222-229,
1968.
Letnansky, K. Strain Variations in the Utilization of Hexose by
Ehrlich Ascites Tumor Cells. Biochim. Biophys. Acta, 165: 364-373,
1968.
Letnansky, K., and KIc, G. M. Glycerophosphate Oxidoreductases
and the Glycerophosphate Cycle in Ehrlich Ascites Tumor Cells.
Arch. Biochem. Biophys., 130: 218—226,1969.
Longshaw, I. D., Bowen, N. L., and Pogson, C. I. The Pathway of
Gluconeogenesis in Guinea Pig Kidney. European J. Biochem., 25:
366-371,1972.
Rognstad, R., and Katz, J. Gluconeogenesis in the Kidney Cortex.
Biochem. J., 116: 483-492, 1970.
Safer, B., Smith, C. M., and Williamson, J. R. Control of the Trans
port of Reducing Equivalents across the Mitochondrial Membrane in
Perfused Rat Heart. J. Mol. Cellular Cardiol., 2: I I I - I 24, 197I.
Shonk, G. E., and Boxer, G. E. Enzyme Patterns in Human Tissues.
Methods for Determination of Glycolytic Enzymes. Cancer Res., 24:
709-721,
1964.
16. Shulman, M., and Wood, H. G. Determination and Degradation of
Microquantities of Acetate. Anal. Biochem., 39: 505-520, 1971.
17. Terranova, T., Baldi, S., and Dionisi, 0. Further Observations on the
Glucose-induced Respiration in Ehrlich Ascites Cells. Arch. Biochem.
Biophys., 130: 594-603, 1969.
18. Terranova, T., Galeotti, T., Baldi, S., and Neri, G. Kinetics of Oxygen
Uptake in Ehrlich Ascites Tumor Cells Treated with Rotenone plus
Glucose. Biochem. Z., 346: 439-445, 1967.
19. Umbreit, W. W., Burns, R. H. and Stauffer, J. F. Manometric Tech
niques,
Ed. 4. Minneapolis,
Minn.:
Burgess
Publishing
Co.,
1964.
20. Weinhouse, S. Oxidative Metabolism of Neoplastic Tissues. Advan.
Cancer Res., 3: 1958.
21. Williamson, J. R., Jacob, A., and Refino, C. Control of the Removal
of Reducing Equivalents from the Cytosol in Perfused Rat Liver. J.
Biol. Chem., 246: 7632-7641, 1971.
APRIL 1974
Downloaded from cancerres.aacrjournals.org on April 30, 2017. © 1974 American Association for Cancer Research.
877
Lactate and Pyruvate Metabolism and Reducing Equivalent
Transfer in Ehrlich Ascites Tumor
Joseph Katz, Karl Brand, Sybil Golden, et al.
Cancer Res 1974;34:872-877.
Updated version
E-mail alerts
Reprints and
Subscriptions
Permissions
Access the most recent version of this article at:
http://cancerres.aacrjournals.org/content/34/4/872
Sign up to receive free email-alerts related to this article or journal.
To order reprints of this article or to subscribe to the journal, contact the AACR Publications
Department at [email protected].
To request permission to re-use all or part of this article, contact the AACR Publications
Department at [email protected].
Downloaded from cancerres.aacrjournals.org on April 30, 2017. © 1974 American Association for Cancer Research.