The Effect of Transplantable Tumors on the Seromucoid
Fraction of Rat Serum*
ROBERT
A.
L.
MACBETH
(Department of Surgery and Surgical-Medical
AND
J.
Research Institute,
GEORGE
BE@cIl@sI
University of Alberta, Edmonton,
Canada)
SUMMARY
The seromucoid fraction of rat serum has been investigated by means of serial
analyses of its protein,
mals
subjected
hexosamine,
to various
hexose, sialic acid, and fucose content
experimental
conditions.
Twenty-two
animals
in 168 ani
served
as
normal controls, and the four test groups included : (a) rats bearing Walker 256 car
cinoma, (b) rats bearing Flexner-Jobling
carcinoma, (c) rats given a single intra
muscular injection of denatured Walker tumor, and (d) rats given a single intramus
cular injection of viable rat liver cells. Characterization of the seromucoid fraction
was also attempted by various means.
The data presented indicate that all seromucoid components measured show a
progressive
and marked
increase in Walker
carcinoma-bearing
rats, a pattern
which
is at variance with that observed in Flexner-Jobling carcinoma-bearing rats and rats
receiving a single intramuscular
injection
of either heat-denatured
Walker tumor cells
or viable rat liver cells. The seromucoid produced in response to aggressive tumor
growth is biochemically unlike normal seromucoid in its carbohydrate
composition,
and ultracentnifugation
studies suggest that it contains a seromucoid fraction that
either is not present in the serum of normal rats or is present in quantities that are
incapable of measurement by the methods available.
Interest
crease
in serum glycoproteins
in recent
years
as a result
has shown a marked in
incorporation
of the demonstration
consistently
of
their deviation from normal in a variety of pathological
states.
As a result, considerable progress has been made
toward the clarification of their structure, biosynthesis,
and biological significance (27, 28, 38). Studies on the
normal biosynthesis of the serum glycoproteins utilizing
glucose-C'4 in the rabbit (6) and the rat (26) and glucos
amine-C14 in the latter species (24) would indicate that the
labeled carbohydrates are rapidly incorporated into the
protein-bound hexosamine of the liver and subsequently
into that of the serum glycoproteins.
It has also been
observed (24) that the rate of incorporation
into the
carbohydrate
portion of the seromucoid fraction is many
times faster than into the nonseromucoid fraction of the
plasma proteins.
The incorporation
of S35-labeled DL
methionine into the seromucoid fraction of dog serum has
also been studied before and after total hepatectomy
(14). These studies indicate that the rate of incorporation
is reduced to one-third by hepatectomy and are interpreted
to indicate that the major synthesis of seromucoid occurs
in the liver. In addition, studies on the isolated perfused
rat liver have provided direct evidence for the hepatic
synthesis
* This
of seromucoid
investigation
tional Cancer Institute
was
and
supported
of Canada
confirmed
by
that
a
grant
(NCI * 226).
Received for publication June 15, 1964.
the
from
rate
the
of
Na
of glucose-C'4
into the seromucoid
fraction
is
more rapid than into the other serum glyco
protein fractions (21, 22). There is also evidence to
suggest that in tumor-bearing animals the liver plays a
significant role in the initiation of the elevated serum
glycoprotein levels observed under these circumstances
(17).
In the human also, supporting evidence for the hepatic
biosynthesis
of serum
glycoproteins
has been reported.
Biological half-life studies on I'31-labeled orosomucoid, for
example, have indicated that the rate of synthesis of this
compound is increased in patients with various inflam
matory diseases but decreased in those with parenchymous
liver disease (35).
It is now well established that certain disease states are
characterized by elevation of the total serum glycoprotein
levels. Investigations
in various laboratories have con
firmed such elevation in patients suffering from cancer (1,
12, 16, 23, 24, 28, 38) and in animals
bearing transplantable
tumors (3, 8, 18, 25, 34). More recently attention has
been centered on the seromucoid fraction of total serum
proteins, since this fraction has been demonstrated
to be
particularly rich in protein-bound
carbohydrates
(36) and
to be the major contributor to the glycoprotein elevation
observed in transplantable malignant tumors in the rat
(34). Seromucoid has been defined as a group of macro
molecular
conjugated
serum proteins
that
are soluble in
2044
Downloaded from cancerres.aacrjournals.org on June 15, 2017. © 1964 American Association for Cancer Research.
MACBETH
AND
BEiu@sI—Seromucoid
0.6 N perchlor]c acid but insoluble in 1 Mphosphotungstic
acid (7, 36). The demonstration
that certain disease
processes have a marked effect on seromucoid levels has
given rise to the suggestion that seromucoid may even have
a different site of origin as compared with the other serum
glycoproteins and may respond in a different manner to
appropriate stimuli (21).
Since there appears to be a paucity of information
regarding changes in the seromucoid in animals bearing
transplantable
tumors (3, 34) it was decided to initiate a
study of this specific glycoprotein complex in such a situa
tion. Serial determinations were carried out in rats bear
ing intramuscular Walker 256 carcinoma and in rats bear
ing subdermal
Flexner-Jobling
carcinoma. Control
groups consisted of rats given inoculations intramuscularly
of heat-denatured Walker 256 carcinoma or viable rat
liver cells. Seromucoid was studied by estimating the
protein, hexose (galactose and mannose), hexosamine
Adult
of Rat
2045
Serum
Subsequently 0.3 ml. of this homogenate was injected into
the thigh musculature of the right hind limb of the recip
ient rats by means of a 1-cc. tuberculin
syringe fitted with
a 23-gauge needle.
Test animals were killed in groups commencing 48
hours following implantation
by exsanguination
from the
abdominal aorta under light ether anesthesia.
The blood
samples were suspended for an hour at room temperature,
then
centrifuged
for
15 minutes
in an
International
centrifuge following which the sera were collected.
Isolation of seromucoid was carried out by the method of
Weimer and Moshin (33). However, in order to provide
for duplicate analyses of each color reaction, it was neces
sary to combine the serum from two animals
before bio
acid), and fucose in this serum protein fraction.
chemical analysis. The total serum protein and sero
mucoid protein were determined by the method of Wolfson
and Cohn (40). Estimation of seromucoid hexose
(galactose and mannose) was carried out by the colon
metric method of Lustig and Langer (15) as modified by
Weimer and Moshin (33). Senomucoid fucose was
MATERIALS
assayed by the method of Dische and Shettles (9).
Estimation of the seromucoid hexosamine (glucosamine
(glucosamine
neuraminic
Fraction
and
galactosamine),
sialic
AND
Sprague-Dawley
(N-acetyl
METHODS
rats weighing
gm. were selected for this study.
per cage and maintained
acid
from 240 to 310
They were housed two
on a standard
grain diet (13) with
tap water ad libitum. In Gioup 1, 22 rats were selected
as untreated controls and were killed in four separate
groups.
In Group 2, 35 rats received intramuscular
implantations
of Walker 256 carcinoma by the technic
previously described (18). In Group 3, 33 rats were
and galactosamine) was performed by hydrolyzing the
seromucoid sample with 3 N HC1 at 100°C. for 4 hours in
sealed
serum
bottles.
The
hydrolytic
product
thus
obtained was purified on a Dowex 50-X8, 200-400 mesh
resin column (5). The effluent thus obtained was analyzed
for hexosamine by the colorimetnic method of Elson and
Morgan
(10).
The isolation and purification of seromucoid sialic acid
given subcutaneous implants in the right axilary region of
Flexner-Jobling carcinoma (29).
In Group 4, 39 rats received a single intramuscular
injection of heat-denatured Walker 256 carcinoma. The
were achieved by the technic of Svennerholm
(30).
After
technic employed for this 4th group was as follows. A
Sprague-Dawley
rat bearing a 5-day-old Walker tumor was
anesthetized with ether, and the entire tumor was removed
To obtain more precise characterization of the sero
mucoid fraction under normal and pathological conditions
purification on a Dowex 2-X8, 200-400 mesh resin column
the effluent was assayed for sialic acid content
ren's T.B.A. color reaction (32).
by War
Three parts of sterile saline were added, and the mixture
it was subjected to further analysis in normal rats, rats
bearing 14-day-old Flexner-Jobling
carcinoma, and rats
bearing 14-day-old Walker 256 carcinoma.
Seromucoid
was isolated from the serum by the method of Winzler
was gently homogenized for 60—90seconds.
The homog
enate was then transferred
to a 25-mi. test tube and
heated in an 85°C. water bath for 2 hours, following which
were then dialyzed against running distilled water for 24
hours and freeze-dried.
The seromucoid fractions thus
procedure it was cooled to room temperature and tested for
its capacity to carry out oxygen consumption and anaer
obic glycolysis (31). Since the heat-denatured Walker 256
obtained were finally dissolved in 0.14 N sodium chloride to
give a 1 per cent solution and subjected to analysis in a
Spinco E model Ultracentrifuge at 21°C. Sedimentation
carcinoma homogenate did not exhibit any metabolism by
either method it was assumed that the cancer cells were
devitalized by the heat treatment.
Subsequently 0.3 ml.
patterns were obtained at 4-minute intervals from 0 to 36
minutes after reaching 59,780 r.p.m.
In addition to ultracentnifugation
the seromucoid from
the same three experimental groups was subjected to im
intact and dissected free of connective tissue.
tumor
tissue were transferred
of the heat-denatured
Blocks of
into a glass homogenizer.
homogenate was injected into the
thigh musculature of the right hind limb of the recipient
rats by means of a 1-cc. tuberculin syringe fitted with a 23gauge needle.
In Group 5, 39 rats received a single intramuscular
injection of viable rat liver cell homogenate.
The homog
enate was prepared in the following manner. The liver
was removed from the donor Sprague-Dawley rat and
placed in a beaker containing cold normal saline. A
piece of liver tissue was then transferred to a glass homog
enizer, 3 volumes of sterile saline were added, and the
mixture
was
homogenized
gently
for
60—90 seconds.
(36, 39).
The perchioric acid-soluble protein samples
munoelectrophoresis and to paper and starch gel electro
phoresis with a Tnis-EDTA-borate buffer at pH 8.9 (2), a
veronal buffer at pH 8.6, and a citric acid-disodium
phos
phate buffer at pH 4.5 (19).
RESULTS
Ra13 bearing Walker 256 carcinoma.—The results ob
tamed in the rats bearing Walker 256 carcinoma are
recorded in Table 1. It can readily be seen that the sero
mucoid proteins manifested a continuous rise following
transplantation
which reached a maxinum of 966 mg. per
Downloaded from cancerres.aacrjournals.org on June 15, 2017. © 1964 American Association for Cancer Research.
2046
Cancer Research
Vol. 24, December 1964
TABLE 1
EFFECT
OF GROWTH
OF INTRAMUSCULAR
WALKER
256
CARCINOMA
ON SEROMUCOID
CENT
SEIOMU
TUMOR
PLASMA
PROTEIN
(GM.
AGE
corn pio
PROTEIN
(xo.
TEIN
PER CENT)Pai
(DAYs)“3TOTALPER CENT)*SEROMIJCOID
@
:@HExosEs*tHExosAinNEs*tSIALIC
F/HACon
± 0.04
trol
2
4
7
4
7
4
*
± 41
5.97 ± 0.06 326.0 ±16.0
5.46
9.25
5.18 ± 0.20 742.8± 43*
14.34
4
Including
±0.10 743.0
±0.32 966.0
±0.50 875.2
±0.40282.0
610.0
the
standard
± 1.13
i/HA
± 0.15
1.08 0.58 0.041
0.99 0.59 0.061
9.91 ± 0.10
0.70 ±0.02
13.10 ± 0.35@ 1.36 ± 0.01
25.90± 3.10@ 38.60 ± 5.40@ 21.80 ± 1.19* 1.95± 0.01@
29.60 ±1.90@ 55.70 ±5.50@ 38.90 ±4.60@ 2.17 ± 0.38@
44.80 ±3.20@ 59.80 ±5.80k 39.40 ±5.10@ 3.42 ± 0.29@
18.44 ± 0.76*
22.12 ± 1.48@
17.04 ± 0.94k
22.35 ± 1.75*
0.67 0.56 0.051
±26@ 14.10
0.53 0.7 0.03@
±110@ 17.76
0.75 0.66 0.057
±92* 17.86 39.40 ± 5.l0@ 59.30 ±7.30k 45.45 X 3.50@ 2.35 ± 0.21k 0.66 0.77 0.040
12.4212.90
±[email protected]
54.45 ±7.90k9.28
0.025
33.50 ±[email protected]
35.46 ± [email protected] ±0.200.96 0.620.69
0.650.084
deviation.
t Seromucoid protein-bound
@
± 1.80
± 0.97
5.65 ±0.15 522.8 ±68@
5.27
5 5.44
13
5 4.9
16
4.91
2022 66.08
10
ACIDtFucosE*tRATIOS
IN
TOTAL
carbohydrate in mg/100 ml serum.
H, hexoses; HA, hexosamines; SA, sialic acid; F, fucose.
§
P = <0.05.
cent by the 13th day, following which it decreased.
Sero
mucoid normally constituted 4.64 per cent of the total
serum protein of the rat, but at its peak elevation fully
17.86 per cent of the total serum protein is contributed by
the seromucoid fraction.
Significant elevation of the seromucoid-bound
hexose
and hexosamine were recorded by the 2d day following
tumor implantation,
whereas significant
elevations
of
sialic acid (N-acetylneuramiic
acid) and fucose first be
came manifest on the 4th and 7th days, respectively.
The
maximum elevations of the hexoses, hexosamines,
and
fucose occurred on the 13th day, and that for sialic acid
occurred on the 16th day. In the case of all seromucoid
bound monosaccharides
the maxima were followed by
progressively decreasing levels.
One might be tempted to suggest that the elevation of
the seromucoid-bound
monosaccharides
merely reflects an
increase in normally constituted
seromucoid, but such
does not appear to be the case. In Table 2 the various
seromucoid-bound monosaccharides are expressed relative
to the seromucoid protein. It is readily seen that, al
though the hexose content of seromucoid remains relatively
TABLE
EFFECT
OF GROWTH
the study, it is relatively rich in
and sialic acid on and after
the 10th post
transplant day. The concentration of fucose in sero
mucoid, on the other hand, appears to be reduced through
out virtually the entire period of observation.
Since there is now considerable evidence that the hexos
amine molecule constitutes the binding link between the
oligosaccharide
and the polypeptide
chain (27) it is of
interest to review the ratios of the various seromucoid
bound monosaccharides relative to hexosamine following
transplantation
(Table 1). Although the sialic acid:
hexosamine ratio remains relatively constant
period of observation, the fucose:hexosamine
during the
ratio drops
to about one-half its normal value by the 2d post-trans
plant
day,
and
that
of hexose-hexosamine
reaches
a
relatively stable state at two-thirds of normal by the 7th
post-transplant
day.
Rats bearing Flexner-Jobling
carcinoma.—The results
obtained in rats bearing Flexner-Jobling
carcinoma are
summarized
in Table 3. However,
before they are
presented it appears desirable to detail the natural history
of this tumor as observed in our laboratory.
This slowly
growing tumor is well established only by the 7th day
following transplantation,
by which time it has attained a
2
OF INTRAMUSCULAR
constant throughout
hexosamine
WALKER
256 CARCINOMA
ON PROTEIN-BOUND CARBOHYDRATES RELATIVE TO SEROMUCOID
diameter of 5—10mm.
consistently
observed
During the first 9 days it was
to be limited
to the subdermal
adipose tissue, but after that time varying degrees of
contact with and attachment to the subadjacent muscle
acidFucose*Control4.64.73.30.4025.65.23.00.2144.24.22.50.2673.55.22.90.26104.07.55.20.29134.66.24.10.35164.56.85.20.27205.58.95.80.22
Tumor
age (days)HexosesHezosaminesSialic
were apparent. If permitted to run its natural course the
Flexner-Jobling carcinoma led to death of the host in from
45 to 50 days. However, it is noteworthy that in our
experience it remained limited to the pectoral region and
that lymph node and distant metastatic involvement did
not occur.
The effects of the growth of Flexner-Jobling carcinoma
on the seromucoid fraction, as recorded in Table 3, are in
sharp contrast
S Protein-bound
mucoid
protein.
carbohydrates
expressed
in
mg/100
mg
sero
to those observed
with the Walker tumor.
Significant elevation of the seromucoid protein does be
come apparent by the 9th post-transplant day but is
Downloaded from cancerres.aacrjournals.org on June 15, 2017. © 1964 American Association for Cancer Research.
MACBETH
minimal
compared
AND
BEKEsI—Seromucoid
with that seen in Walker tumor-bear
ing rats. Changes in the seromucoid-bound monosac
charides are also late in their occurrence and, although
significant, are much reduced in degree.
In the case of
sialic acid no significant deviation from normal occurred.
The ratios of the various monosaccharides
to hexosamine
are also included for comparison with Table 1.
Rats given intramuscular
injections of heat-denatured
Wa1ker 256 carcinoma.—The results of this phase of the
experiment are presented in Table 4. It may be seen that
statistically
significant elevations above normal occurred
in relation to the seromucoid protein and the seromucoid
bound hexosamine between the 2d and 7th day only and
that the sole other seromucoid-bound
monosaccharide
to
manifest significant elevation was the sialic acid and then
only on the 2d day.
Rats given intramuscular injections of viable rat liver cell
homogenate.—The results in relation to this 4th group are
presented in Table 5. It may readily be seen that the
only significant deviation from normal occurred on the 2d
Fraction
of Rat
2047
Serum
day and was manifest in relation to the seromucoid protein
and seromucoid-bound hexosamine.
Characterization
of seromucoid.—In
the course
of a pre
liminary investigation a number of preparations of sero
mucoid from normal rats and rats bearing Flexner-Jobling
and Walker
starch
256 carcinoma
gel electrophoresis
were subjected
and
to paper and
to immunoelectrophoresis,
but in all instances the movement of the seromucoid from
normal and from tumor-bearing
rats was similar and cor
responded to that of a1 globulin.
Representative
sedimentation
patterns obtained on a
Spinco E model ultracentrifuge
are reproduced in Plate 1.
It is apparent that the seromucoid fraction from normal
rats gives a single peak, with a sedimentation
constant of
3.5S. Preparations
from rats bearing Flexner-Jobling
carcinoma
contain an almost identical
component
with a
sedimentation
constant
of 3.57S and, in addition,
a fast
moving fraction the S@o‘a'
of which could not be determined
owing to the small amount present.
The sedimentation
pattern
of seromucoid from rats bearing Walker 256
TABLE 3
EFFECT
OF GROWTH
OF SUBDERMAL
FLEXNER-JOBLING
CARCINOMA
ON SEROMUCOID
CENT
@
SEROIIUCOID
TUMOR
AGE
(DAYS)No.
ANI
PROTEIN
MAI.STOTAL PER
(GM.
PROTEIN
(MG.
PROTEIN
F/HAControl
ACIDtFUCOSEtRATIOS
H/HA SA/HA
PROTEINHaxosEs*tHEXOSAILINES'tSi@LIc
± 41
± 0.04
@
IN
CENT)*SEROMUCOID
p@ CENT)@PER
± 0.97
± 1.13
± 1.80
2
6
6.21 ±0.50 211 ±29
3.40
13.35 ±0.43
4
4
6.2 ± 0.29 236 ± 9
3.81
7
4
5.9
227 ±15
3.85
11.01 ± 0.70
12.72 ± 0.95
13.25± 1.95 14.01± 0.33 10.40 ± 0.20
0.60
Including
the
standard
5.47
± 0.15
0.89 ±0.05
1.30
0.81 0.086
0.86 ± 0.01
0.76
0.50 0.060
0.87 ±0.08
0.68
0.39 0.050
1.16± 0.03 0.95 0.74 0.083
17.94 ±2.70@ 27.07 ±3.6$ 12.50 ± 2.30 2.46 ± 0.19$ 0.66 0.46 0.091
6 6.31 ±0.29 449 ±26@ 7.12
13
4.58
13.40 ± 1.60 25.63± 5.7$ 7.24 ± 2.10 1.35± 0.01 0.52 0.28 0.053
4
6.63 ± 0.26303.5 ± 31
17
401 ± 12$4.64
6.0412.90
14.90± 0.3613.40
26.92± 1.15$9.28
9.81 ± 1.101.12
1.43 ± 0.270.96
6.64 ± 0.10282
0.053a46.08
0.550.69
0.360.084
2622
5
9
6.34 ± 0.08 347 ± 15*
10.26 ±0.48
8.26 ±1.80
14.40 ±0.50
7.21 ±0.46
18.60 ±2.10@ 7.32 ±1.25
deviation.
t Seromucoid protein-bound carbohydrate in mg/100 ml serum.
@
H, hexoses;HA,hexosamines;SA,sialicacid; F, fucose.
$P = <0.05.
TABLE 4
SEROMUCOIDTUMOR
EFFECT
OF INTRAMUSCULAR
F/HAControl
OF HEAT-DENATURED
WALKER
256
CARCINOMA
ON
CENT
SEROMUCOID
PLASMA
AGE
(DAYS)No.
INJECTION
PROTEIN
(Mo.
PROTEIN
(GM.
ANI
PER CENT)PER
MALSTo@iu. PER CENT)SEROMUCOID
PROTEININ
ACIDtFUCOSEtRATIOS
H/HA
SA/HA
PROTEINHExOSE5'tHEXOSAMINES'tSIALIC
@
0.15
± 1.13
± 1.80
± 0.97
13.60 ± 1.80 23.40 ± 2.04$ 15.83 ± 0.70$ 1.57± 0.24 0.58 0.68 0.067
6.62
11.59± 2.23 19.28± 1.46$ 9.68 ± 1.16 1.19± 0.20 0.6 0.5 0.062
6.20 ± 0.10 367 ± 10.9$ 5.92
6
15.11± 0.96 22.26± 0.98$ 7.86 ± 0.44 1.59± 0.22 0.68 0.35 0.071
6.16 ± 0.15 379 ± 20.5$ 6.15
6
12.14± 1.06 12.47± 2.07 6.57 ± 0.33 1.12± 0.04 0.97 0.53 0.090
4.56
5
6.09 ± 0.21 278 ± 23.0
13.77± 0.37 12.59± 0.45 6.14 ± 0.19 1.73± 0.05 1.09 0.49 0.137
3.91
6.42 ± 0.30 251 ± 49.0
5
6.72 ± 0.80 0.88 ± 0.01 0.97 0.64 0.083
10.20 ± 1.70 10.55 ± 1.30
3.02
6 6.72 ±0.90 203 ± 12.6
0.75 ± 0.030.96
0.690.69
0.057
9.00 ± 0.4113.40
13.05± 1.159.28
8.20 ± 0.151.12
0.630.084
3.7412.90
250 ± 10.04.64
56.08
6.69 ± 0.26282
± 0.04
2
6
4
7
10
13
16
2022
*
Including
± 41.4
5.83 ±0.27 386 ± 29.9$
the
standard
deviation.
t Seromucoid protein-bound carbohydrate in mg/100 ml serum.
@
H, hexoses; HA, hexosamines; SA, sialic acid; F, fucose.
§
P = <0.05.
Downloaded from cancerres.aacrjournals.org on June 15, 2017. © 1964 American Association for Cancer Research.
Vol. 24, December 1964
Cancer Research
2048
TABLE
EFFECT
OF INTRAMUSCULAR
INJECTION
5
OF VIABLE
LIVER
CELLS
ON SEROMUCOID
CENT
@
@
TUMOR
AGE
(DAYS)No.
F/HAControl
2
4
7
16
2022
ACIDtFUCOSE@fRATIOS
SA/HA
PROTEINHExoszs*tHEXOSAMINESfSIAUc
± 1.80
± 1.13
± 0.15
23.50± 2.90$12.12± 1.11 1.17± 0.09 0.67 0.52 0.050
12.89± 2.60 8.82 ± 1.30 0.93 ± 0.10 0.97 0.68 0.072
17.15 ± 6.50 10.80 ±1.20 0.82 ± 0.08 0.8
4.34
0.63 0.048
284 ± 47
6.55 ± 0.30
6
15.00 ± 1.60 7.10 ± 0.45 0.83 ± 0.10 0.97 0.47 0.055
4.20
6.03 ± 0.18 253 ± 23
6
12.41 ± 2.20 6.70 ± 0.23 1.09 ± 0.07 0.97 0.54 0.088
4.29
270 ± 24
6.29 ± 0.30
6
11.33 ±1.48 14.46 ± 0.20 7.47 ± 0.15 0.88 ± 0.03 0.78 0.52 0.061
4 6.5 ±0.10 242.5 ± 1.5
3.73
16.67 ± 2.079.289.06 ± 0.971.121.00 ± 0.020.960.970.69
0.540.084
0.060
66.086.35 ±0.50282 288 ± 6.44.64 4.5412.90 16.15 ±0.5213.40
7
4
10
13
IN
PLASMA
PROTEIN
(M?.
PROTEIN
(Gie.
AN!PER CENT)PER
MALSTOTAL PER CENT)SEROM@UCOm
a Including
± 41
364 ± 57$
270 ± 19
± 0.04
6.18 ± 0.20
6.50 ± 0.10
the
standard
5.89
4.15
± 0.97
15.79± 1.28
12.51± 1.08
13.76 ± 4.60
14.58± 2.70
12.01 ±0.59
deviation.
t Seromucoidprotein-bound carbohydrate in mg/100ml serum.
@
H, hexoses; HA, hexosamines; SA, sialic acid; F, fucose.
§
P = <0.05.
carcinoma was found to be different, however, from that in
either of the previous two groups. Two peaks were
clearly apparent—one consisting of a slowly moving com
ponent (4.46S) and the other a fast-moving component
(23.55).
DISCUSSION
On the basis of these data it would appear reasonable
to
conclude that although nonspecific stimulation (denatured
tumor cell or viable liver cell injection) evokes a mild
transient elevation of the seromucoid protein and sero
mucoid-bound
hexosamine in the rat this response is in
sharp distinction to the marked and prolonged response
observed with aggressive tumor (Walker 256 carcinoma)
growth which affects not only the seromucoid protein but
all of the seromucoid-bound
monosaccharides
as well.
ACKNOWLEDGMENTS
The authors would like to express their appreciation to Miss
S. Bice and Mr. E. Sugden for their valuable technical assistance
in
rats
bearing
Walker
256
carcinoma
reveals
that
the
relative proportions of seromucoid-bound
hexose, hexos
amine, fucose, and sialic acid differ significantly from those
observed in normal animals.
This observation
would
appear to be further substantiated
by the ultracentrifuga
well established Walker 256 carcinoma contains a sero
investigation
further
will, however,
characterize
be necessary
this seromucoid
to isolate
fraction.
PLATE 1.—Sedimentation
and
this
study.
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2049
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The Effect of Transplantable Tumors on the Seromucoid
Fraction of Rat Serum
Robert A. L. Macbeth and J. George Bekesi
Cancer Res 1964;24:2044-2051.
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