Download Gene Activation of Molecules with

[CANCER RESEARCH 36, 3503-3509, September i976]
Gene Activation of Molecules with Carcinoembryonic Antigen
Determinants in Fetal Development and in Adenocarcinoma of the
Colon1
Allyn H. Rule2 and Mary E. Kirch
Graduate Department of Biology, Boston 02167, and Department of Dermatology and Medicine, Tufts University School of Medicine, Bostor@,Massachusetts
02111
Summary
“Fingerprints―
of 0.9% NaCI solution extracts obtained
from fetal guts and individual adenocarcinoma of the colon
show a randomized pattern of expression of carcinoem
bryonic antigen (CEA) determinants by CEA radioimmu
noassay and isoelectric focusing. All CEA-containing anti
gens found in a pool of 20 primary adenomas were found at
some stage in fetal development. No single CEA-reacting
peak was typical of any one period of fetal development.
When fetal gut profiles were grouped according to trimester
in utero, however, an expanded gene pool was found in the
second trimester which correlates well with maximum gas
trointestinal growth and differentiation. Isoelectric focus
ing-CEA radioimmunoassay profiles of individual primary
adenomas were similar to but never identical with individual
fetal gut profiles. “Fingerprints'
‘
of metastatic adenomas of
entodermal origin showed quantitative and qualitative in
creases in molecules with CEA determinants unlike these
latter categories. Such data suggest that both integrator
and controller gene activities may be lost in metastatic
disease. Rather than “phase-specific
gene sets―on differ
ent chromosomes being activated by various oncogenic
modalities, it is more probable that individual chromosomes
are involved in oncogenesis. While more data are needed to
confirm this idea, it is safe to say that the expression of
molecules with CEA determinants need not be caused by
either demepressive or meexpressive gene activation. These
data point to the individuality of gene expression of mole
cuies with CEA determinants both in fetal development and
in early neoplasia. Since CEA-reacting molecules were not
found in tumors of ectodemmal or mesodermal origin by
these methods, such products should be termed carcino
developmental antigens of entodemmal or colonic origin.
Analyses of nonrepetitive sequences of mRNA and DNA at
various developmental stages have been used to construct
models of gene activation during mammalian embryogene
sis (12). Current data from several laboratories suggest that
I Presented
at
the
Symposium
“Cancer
and
Chemistry―
as
part
of
the
Fourth Conference on Embryonic and Fetal Antigens in Cancer, November 2
to 5, 1975, Charleston, S. C. This investigation was supported by Grant CA
12924-04, awarded by The National Cancer Institute, Department of Health,
Education, and Welfare.
2 Presenter.
partially overlapping phase-specific genes on multiple chro
mosomes are expressed in a limited manner during any 1
period of differentiation (30). Quantitative and/or qualitative
control of gene expression may be thought to consist of a
delicate interplay of sensor, integrator, activator, receptor,
and producer genes or gene products which can react at
transcriptional , pretmanslational , translational , and post
translational levels for any 1 set of genes during develop
ment (4, ii).
Our own laboratory has been studying models of gene
activation during embryogenesis, using final gene products
present in both adenocarcinoma of the colon and fetal gut
which have CEA3 determinants. Since final gene products
may be potentially modified at several different levels, this
paper will attempt to make simple although preliminary
analogies in comparing normal fetal, adult, and neoplastic
gene patterns. However, normal colon products with CEA
determinants can most definitely be “fingerprinted―
and
compared to normal gut differentiation. The concept that
multiple CEA determinants are found on many molecules of
entodermal origin has been amply verified by many laboma
tories, using a wide spectrum of currently available tech
niques (Table 1).
While some investigators suggest that such data indicate
heterogeneity of CEA-reacting molecules (3, 7) or the possi
bility of isomenic forms (35), the sharp isoelectnic focusing
pattern of a 0.9% NaCI solution extract of a fetal gut at 21
weeks shown in Chart 1 clearly indicates that different ge
netic forms of molecules containing CEA determinants can
be sharply isolated and identified without degradation. Urea
isoelectnic focusing of tumors of mesodermal or ectodermai
origins was found no longer to express CEA reactivity,
even though crude 0.9% NaC1solution tumor homogenates
on plasma from these patients were initially CEA positive by
CEA radioimmunoassay. Our studies are thus confined to
molecules with CEA determinants of entodermal origin.
Initially, 2 molecules containing CEA determinants were
thought to represent reexpressed early developmental gene
products of colonic origin. Indeed the original term “carci
noembryonic―reflects this early hypothesis (18, 19). Other
data suggested that tumors may also produce excessive
quantities of so-called normal CEA-reacting autoantigens
which, once expressed in development, may normally be
3The abbreviation used is: CEA, carcinoembryonic antigen.
SEPTEMBER1976
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3503
A. H. Rule and M. E. Kirch
Table 1
Evidence for different molecules with CEA determinants
1 . Precipitation
in Ouchterlony
gel (5, 6, 27, 29, 31 , 34, 46).
2. Uitracentnifugation:molecularweight determinations(8, 24, 45).
3, Affinity chromatography(9, 16, 22, 36).
4, isoelectric focusing (7, 38-40).
5. Localization on colon cells and in meconium
(13, 14, 20, 37, 40).
6. Clinical studies: lack of CEAspecificity and sensitivity for diag
nostic purposes(1, 10 15 21, 23 26 28 32 33 42-44 47 48).
overlapping gene sets occur during gut embryogenesis and
thus probably occur in neopiasia also. Individual and
pooled data of both fetal colon and adenocarcinoma of the
gastrointestinal tract are thus discussed relative to our early
hypotheses of gene action in neoplasia and to newer theo
mies. Additionally, the relationship of gene products with
CEA determinants to normal gut development is shown.
Materials and Methods
1.2 -
. 12
21 Weeks
. II
I
1.0 -
I
I
•
10
‘9
I
0.8-
I
I
‘8
I
Id
I
@
I
I
D
I-'
Id
C.)
I I
@0.6-
I
I
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I
@
I
3'
I
5
0.4-
4
I—
.
0.2 •
3
2
@1
@
An
5
10
___1_
5 20
25
30
35
@+I I
40 45
TUBENUMBER
Chart 1. “Fingerprint―
of molecules with CEA determinants in a 21-week
fetal gut sample. A single peak at pH 4.0 appears with no degradation forms
after isoelectric focusing in 7 M urea with ampholines in the range of pH 2.5
to 10.0.
found in low quantities in adult colonic mucosa (5, 27, 29,
31 ,38, 40, 41).
Utilization of a radioimmunoassay for CEA determinants,
in conjunction with urea-sucrose isoelectric focusing of
adenocarcinoma of the colon and pancreas, initially gave
credence to both these ideas: that cancinogenesis did pro
duce 2 products of demepressive, dediffenentiating, onco
genic mechanisms but that some of these so-called carci
nodevelopmentai glycoproteins were normal adult products
produced in abnormal quantities via tumorigenic mecha
nisms (39, 40). in addition, our early work with fetal gut
suggested that cancinogenesis might involve phase-specific
antigen expression of an earlier developmental period (41).
Analysis of a larger group of fetal colons in this present
study does not exactly substantiate our original hypothesis,
although these assumptions helped us to formulate the
basic rationale for initiating this study. These hypotheses
are found in Table 2 but can be condensed in the following
manner. The total expression of embryonic and normal
colon products in adenocarcinoma of the colon should be
found in a large tumor pool of adenocarcinomas of the
colon as well as in large numbers of individual normal fetal
and adult colon tissue samples. Phase-specific multiple
3504
Fetal guts were obtained after surgery. Relative age in
utero was established both by crown to rump measurement
and by relative length of the total gut. When possible, intes
tines, meconium, and colon were separated. Each colon
was homogenized in 0.9% NaCI solution 1:2 w/v at 4°and
spun twice at 40,000 x g. Biuret protein content was esti
mated on the final supemnatants against appropriate bovine
serum albumin controls. The fetal gut extracts were stored
in appropriate aliquots of —20°
and once thawed prior to
isoelectnic focusing. Samples were then treated with 7 M
urea at 4°for 24 hr and at 45°for 1 hr prior to isoelectnic
focusing in 1.5% pH 3 to 10 ampholines, containing 0.01 M
NaC1 in a 7 M urea-sucrose 110-mi gradient at 10°(2 to 10
ma) by previously described methods. The use of urea both
to separate out these antigens and to provide a dense
matrix for peak resolution is emphasized. Previous reports
have indicated that “antigen
floating―without such treat
ment yields immeproducible, badly resolved peaks (6, 40).
Carefully excised tumor or normal colon tissues were run in
a manner previously described (41). Up to 20 mg of protein
were loaded on each column prior to focusing. However, all
data were normalized to a fixed protein content so that all
isoelectnic focusing profiles can be related to each other as
well as to our earlier studies.
CEA madioimmunoassays were run in duplicate on came
fully dialyzed isoelectrically focused fractions in the most
linear mangeof the control curve, or suitable concentrations
or dilutions made until the amount of CEA-reacting material
in each tube could be accurately ascertained. Aeagents
used for this zirconyl phosphate gel CEA madioimmunoas
say were the kind gift of The Research Division of Hoff
mann-LaAoche Inc., Nutley, N. J. Absombance at 280 nm
and pH were estimated, using equipment with an error of
±0.05A or pH unit. Since little correlation between CEA
values and A28,estimations of protein content was found,
some of our later profiles do not include this variable. Better
peak resolution is likewise found in our later studies when
Table 2
Basic assumptions
1 . Mammalian embryogenesis
proceeds via production
specific multiple overlapping gene sets.
of phase
2. Expressionof embryonic antigens occurs in carcinogenesis.
3, Large pools of tumors of 1 type should furnish all of the possible
embryonic and adult forms of antigens (gene products).
4. individual embryonic tissues should indicate the order, quantity,
and probability
of phase-specific
overlapping
antigen (gene)
sets.
5, Quantitative expression of gene products or various members of
phase-specific
antigen sets need not be found in the same
chromosome.
CANCER
RESEARCH
VOL. 36
Downloaded from cancerres.aacrjournals.org on August 1, 2017. © 1976 American Association for Cancer Research.
Genes Coding for CEA Determinants in Development
our isoelectnic focusing techniques had become highly sys
temized.
2
Results
I0
“
Fingerprints―
obtained
by
CEA
radioimmunoassay
and
isoeiectnic focusing of fetal gut pH and A2@,
determinations
are found in Chart 2. Not only are very few antigen peaks
seen in the earliest profiles but also the earliest antigens
seen are those with pK of 4.0 and 4.5. Peak fractions vary as
to pK1but are found between pH 2.5 and 5.5 in ii of 12 fetal
guts that were focused. Greater numbers and quantities of
peaks are found during periods of maximum growth and
differentiation, and the total gene pool both quantitatively
and qualitatively seems to contract somewhat after this
period of time (about 20 weeks) (Charts 2 to 7).
This concept is more amply illustrated in Chart 8 where
CEA-meacting fetal colon antigens of different tnimesters are
compared with a tumor pool of 20 primary adenocarcino
mas of the colon and pooled data from 8 normal colon
samples obtained postmortem from patients without cancer
or inflammatory bowel disease. Maximum antigenic expres
sion occurs when the gut is undergoing major diffementiat
ing processes and an enlarged gene pool might be envi
sioned. Since peak CEA-reacting determinants in fetal guts
and the primary tumor pool are not significantly different in
Chart 8, it seems logical to assume that quantitative control
of gene activities may not be lost early in the carcinogenic
process. It appears that early neoplastic steps may resemble
fetal development in many ways.
The absolute quantities of CEA-reacting materials relative
9
8
71
6'
5
4
3
2
Chart 3. “Fingerprint―
of molecules with CEA determinants in a 14.5-week
fetal gut. Major peaks are found at pH 5.0, 6.0, 7.5, and 8.0 ±0.2 pH units.
I0O
11.0
10.0
80
@
60
I
6.0 _
I-
4
Id
C.)
@,40
20
4
8
12 16 20 24 28 32
TUBE NO.
Chart 4. “Fingerprint―
of molecules with CEA determinants in a 15-week
fetal
gut.MajorpeaksarefoundatpH 2.0,2.5,3.5,4.0,4.5,5.0,5.5,6.0,7.0,
and 10.0 ±0.2 pH unit.
30
40
TUBE NUMBER
Chart 2. “Fingerprint―
of molecules with CEA determinants found in a
13.5-week fetal colon sample. Peaks appear at pH 4.0 and 4.5 ±0.2 pH unit.
RIA, radioimmunoassay.
SEPTEMBER
to overall protein content found during fetal gut develop
ment are shown more precisely in Table 3. These are com
pared with normal adult and primary adenocancinomas of
the colon in their relative amounts of molecules with CEA
determinants. Average ng of CEA per mg of protein in the
fetal gut (21.5) is similar to that found in normal colon but
much less than that found in primary adenocarcinoma of
the colon. The fact that almost all of the CEA-reacting
fractions of various pK1found in the tumor pool are likewise
1976
Downloaded from cancerres.aacrjournals.org on August 1, 2017. © 1976 American Association for Cancer Research.
3505
A. H. Rule and M. E. Kirch
pH
pH GRADIENT
GRADIENT
@
2 3
4
7
@—@+
9 +‘9
40
I00
FETAL
FETAL
GUT
GUT-26wk
I6wk
40
I@
I'
3.0
4,
r@)
r,)
a,
,‘t
I―
0
@
@
4
I
!
50
a
4
Id
0
0
i;
I
2.0
II
II
It I
1111
I ‘I
l'4t
4
Id
0
I.0
I
I
‘I
.0
5
20
25
TUBE NUMBER
I-,.,-_j@/q
5
TUBE
15
NUMBER
A@-@
25
20
Chart 5. “Fingerprint―
of molecules with CEA determinants in a 16-week
fetal gut. The pH's of major peaks are 2.0, 4.0, 6.5, 7.5, 8.0, and 8.5 (courtesy
of British Journal of Cancer) (39). AlA, radioimmunoassay.
30
35
Chart 7. “Fingerprint―
of molecules with CEA determinants in a 26-week
fetal
gut.Peaksarefoundat3.0,3.5,4.0,4.2,4.5,5.0,6.0,7.5,and 9.0±0.2
pH unit. AlA, radioimmunoassay.
Table 3
Relationship
CEAdeterminants
of quantitative
amounts
of molecules
with
to various
developmentCEA
stages of gut
ma
dioimmu
noassay(ng/mgPeriod
statusA.
(wk)
protein)
utero13.0
In
major14.0
8.0
defined.14.5
5.9
Gut
Histodifferentiation
and
structures of gut are
5.515.0
17.016.0
Id
@
7@
20
I
6
4
Id
0
19.0
developed;19.5
21.0
pear.21.0
33.0
21 .3
5.0
10.2
Mechanical and chemical func
tions of colon are
fine structural
elements ap
12.5
Major differentiation of gut
17.5
Av.
(meanof
normal gut function
55.021.5
62.526.0
iscomplete.B.
Normal adult
colons).C.
Primary
andcinoma
adenocarpresentIon
of the co5
10
15 20 25 30 35
TUBENUMBER
Chart 6. “Fingerprint―
of molecules with CEA determinants in a 19.5-week
fetal
gut.MajorpeaksarefoundatpH 2.5,3.5,4.5,5.0,5.5,6.0,6.5,7.5,and
9.5 ±0.2 pH unit.
found in the pooled fetal gut data shown in Chart 8 lends
credence to the idea that all of the CEA determinants found
in the tumor pool have some relationship to fetal develop
ment. That these products are of entodermal origin is mdi
3506
8
85.0
Early neopiastic changes
unusual cell types
(mean of 20 primary adeno
carcinomas of the colon).
cated by the negative data shown in Chart 9. Of approxi
mately 10 tumors from various nonentodemmal sources that
have been electrofocused in 7 M urea-sucrose gradients,
none so far have shown CEA-reactive peaks by radioimmu
noassay after this procedure. Since many of these tumors
contained CEA-meacting sites prior to focusing (up to 1 @tg/
mg of protein), one can only assume that these CEA deter
CANCER RESEARCHVOL. 36
Downloaded from cancerres.aacrjournals.org on August 1, 2017. © 1976 American Association for Cancer Research.
Genes Coding for CEA Determinants in Development
organs. In Chart 1OAone sees a profile of a typical primary
minants were formed by fortuitous aggregations subse
quently broken by urea dissolution of both H-bonding and adenocarcinoma of the colon with 2 rather low peaks that
hydrophobic bonding in high-voltage electrical fields. The contain CEA-meacting molecules. in contrast, in Chart lOB
individual molecuiesperse from these tumors did not con
one sees the profile of a primary adenocarcinoma of the
tam CEA determinants. We therefore feel confident that we colon with many peaks of CEA-meacting antigens more typi
are analyzing tissue-specific gene products of entodemmal cal of those found in the 16- to 20-week period in utero. This
origin. Whether or not aggregated forms of molecules ex
tumor did go on to form metastases in the same patient.
Chart 1OC shows the CEA profile of a spleen metastasis
pressing CEA reactivity are possibly related to “symbody―
production is a question open to future investigation (2).
from a patient with diagnosed adenocarcinoma of the co
In our current studies, we compared similarities and dif
Ion. This tumor contained 200-fold more CEA than that
femences in CEA “fingerprints―
found in primary colonic
found in Chart bA but only 10-fold more than that found in
tumors in contrast with those that metastasize to other
Chart lOB. Additionally, one antigenic form, the pH 4.0
CEA-reacting molecule, predominates.
COMPARISON
OFNORMALTOONCOCOLON
ANTIGENS
Discussion
The early work of Gold and Freedman (18, 19, 25) con
tamed the invaluable idea that developmental genes are me
expressed in adenocarcinoma of the colon. Urea isoelectnic
.@
40°r
.2
I
‘;300F
@
i
I
I
4
FETAL
GUT
I
£IJst Trimester
D 2@d
Trimester
— 3
200J
10@
Trimester
-- - meconium
0
8@
V
z
@
&
NORMALCOLON
200
6
(8)
Cancer
ofBreast,
Thymus,Thyroid,
Prostate,
Shin
I00
A AA@AA
A
4+
2
‘,
Chart 8. Comparison of normal fetal colon antigens containing CEA de
terminants by trimester in utero with those found in a pool of 20 primary
adenocarcinomas of the colon and pooled data from 8 normal adult colons.
RIA, radioimmunoassay.
Iv
@U
SQ
4U
TUBENUMBER
Chart 9. Lack of “fingerprints―
found by isoelectric focusing of tumors of
nonentodermal origins in 7 M urea followed by CEA radioimmunoassay (RIA).
of each tube.
C
a
7.0
4,
60
V
x
4
Id
0
a,
C
50
E
4,
0
4
TUBE
NUMBER
TUBE
NUMBER
TUBE
NUMBER
Chart 10. A, “fingerprint―
of primary adenocarcinoma of the colon with peaks at pH 2.5 and 5.0 ±0.2 pH unit. B, “fingerprint―
of a primary
adenocarcinoma of the colon which went on to metastasize in the liver. Peaks are found at pH 2.0, 3.0, 3.5, 4.0, 4.5, 5.0, 6.5, and 9.0 ±0.15 pH unit. C,
“fingerprint―
of a liver metastasis of an adenocarcinoma of the colon. The peak tube occurs at pH 3.0 but between 100 and 200 ng of CEA are found in poorly
resolved peaks between pH 4.0 and 9.0. RIA, radioimmunoassay.
SEPTEMBER
1976
Downloaded from cancerres.aacrjournals.org on August 1, 2017. © 1976 American Association for Cancer Research.
3507
A. H. Rule and M. E. Kirch
focusing in conjunction with CEA-radioimmunoassay in our
selective advantages of individual cancer cells to colonize
at distant points in a different organ millieu.
own laboratory with our initial 2 fetal gut samples not only
supported this contention (40) but indicated some ‘
‘phase Our present hypothesis is that metastic processes in co
overlap―between the 2 fetal gut “fingerprints―
(41). The ion cancer involve loss of integrator and controller genes
data contained in this study on 12 fetal gut samples can no which regulate the spectrum and quantity of molecules with
CEA determinants. Work is currently under way to support
longer support these earlier hypotheses. While we had ex
pected the data to conform to either Model System A or B these preliminary hypotheses.
More clear-cut relationships are found with “fingerprints―
shown in Chart 11, these data clearly indicated that both
fetal gut development and early neoplasia exhibit random
of individual fetal gut extracts to fetal development. These
indicate that a small number of CEA-reacting molecules
ized individual gene product patterns that are not confined
initially are present. Quantitative expression of molecules
to any 1 period of embryonic, fetal, or normal adult develop
with CEA determinants is likewise low at this stage of devel
ment. In eliminating Model A by inference, we indicate that
opment where histodifferentiation and major gut elements
stepwise demepressivedifferentiation of so-called adult cells
via oncogenic mechanisms most probably does not take are being formed. During maximal gut differentiation, the
gene pool expands to synthesize more molecules with CEA
place in adenocamcinoma of the colon. Conversely, em
determinants. At this point fine structural elements of the
bryonal type cells that potentially might be present in small
numbers in any one tissue or organ system need not have gut are being defined, and mechanical and chemical func
carcinogenic potential to allow Model System B to take tions of the colon are in the process of definition. In gen
place. Additionally, phase-specific overlapping gene sets eral, quantitative expression of CEA reactivity is likewise
increased. When major gut differentiation is complete, the
that might infer Model A or B as a basis for fetal antigen
expression are likewise precluded from our individual anal
gene pool contracts slightly, as does the total amount of
CEA reactivity.
yses of 12 fetal guts and more than 12 primary and meta
static adenocarcinomas of the colon.
These relationships are more clearly expressed in Chart 8,
However, pooled data are in accord with the concept that
where data are pooled by trimester in utero and compared
with normal and neoplastic gut activities of molecules with
overlapping phase-specific gene sets are a factor in normal
gut development. The randomized patterns of CEA meactiv CEA determinants. The gene pool starts out small initially,
ity shown both in development
and in primary
adenocarci
expands during the 2nd trimester, and contracts somewhat
noma of the gut may indicate the possibility that only 1 in the last trimester. Such data would not be in disagree
ment with changes in transcription of nonrepeated DNA
carcinogenic factor superimposed on normal gene expres
sion need be present for a switch into a neoplastic pattern
sequences in neonataland fetalmice (17).
This study clearly indicates that radioimmunoassays for
of CEA reactivity.
carcinodevelopmental antigens may be used as delicate
In contrast, metastatic potential seems to infer that, after
probes to investigate normal growth and development of
an initiating oncogenic event takes place, gene interactions
are upset, integrator genes begin to code for a wider array
specific organ systems. The quantitative and qualitative
expression of final gene products might be used as addi
of genes similar to the most active phase of gut diffementia
tional probes to correlate with the production of non repeti
tion, and, finally, controller genes cannot limit production
of membrane-localized molecules with CEA determinants. It tive DNA and mRNA. It is clear that knowledge of both
normal and fetal developmental patterns will be essential to
is most probable that 1 or more of these CEA-reacting
understanding neopiastic growth mechanisms in different
molecules occurring with acidic pK1 allow migration and
organ systems.
MODEL SYSTEMS: Expression of Oncofetal Colon Antigens (genes)
let
2nd
3nd
TRIMESTER TRIMESTER TRIMESTER NEONATAL
ADULT
A STEPWISEDEDIFFERENTIATION
._____
a
t
.
@
t_@
@__e
a
a
a
a
B. STEPWISEDIFFERENTIATION
t
*@
a
a
a
a
a
a
a
a
A
C. RANDOM DIFFERENTIATION
a
( DEDIFFERENTIATION)
&—
*
a
a
t
*
*
a
*
*
a
a
Chart 11. Model systems that potentially may be involved in the expres
sionof developmental
or oncofetalcolonantigens(genes).
3508
1. A Joint National Cancer Institute of Cancer/American Cancer Society
Investigation: A Collaborative Study of a Test for Carcinoembryonic
Antigen (CEA) in the Sera of Patients with Carcinoma of the Colon and
Rectum. Can. Med. Assoc. J., 107: 25-33, 1972.
2. Apffel, C. A., and Peters, J. H. Tumors and Serum Glycoproteins: The
Symbodies. Progr. Exptl. Tumor Res., 12: 1-54, 1969.
3, Banjo, C.. Shuster, J., and Gold, P. Intermolecular Heterogeneity of the
Carcinoembryonic Antigen. Cancer Res., 34: 2114-2121 , 1974.
4. Britten, R. J., and Davidson, E. H. Gene Regulation for Higher Cells: A
Theory. Science, 165: 349-357, 1969.
5. Burtin, P., Martin, E., Sabine, M. C., and von Kleist, S. Immunologic
Study of Polyps of the Colon. J. NatI. Cancer Inst., 48: 25-29, 1971.
6. Carrico, R. J., and Usategui-Gomez. M. The Isolation of Carcinoem
bryonic Antigen from Tumor Tissue at Neutral pH. Cancer Res., 35:29282934, 1975.
t___
a
References
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8. Coligan, J. E., Lautenshleger, J. T., Egan, M. L., and Todd, C. W. Isola
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9, Cooper, A. G., Brown, M. C., Kirch, M. E., and Rule, A. H. Relationship
CANCER RESEARCHVOL. 36
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Genes Coding for CEA Determinants in Development
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15.
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SEPTEMBER1976
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3509
Gene Activation of Molecules with Carcinoembryonic Antigen
Determinants in Fetal Development and in Adenocarcinoma of
the Colon
Allyn H. Rule and Mary E. Kirch
Cancer Res 1976;36:3503-3509.
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