Vol. August
178,
No. 15,
3, 1991
BIOCHEMICAL
BIOPHYSICAL
AND
RESEARCH
COMMUNICATIONS
Pages 1099-1104
1991
ENHANCING EFFECTSOF HUMANMACROPHAGE COLONY-STIMULATING FACTOR ON THE SECRRTION OF HUMANCHORIONICGONADOTROPIN BY HUMANCHORIONICVILLOUS CJILLSAND tPA30-1 CKLLS
Shigeru
1,+
Saito
, Hami Saito' , Kazuo Motoyoshi3 , and Motohiko Ichijo'
IDepartment of Obstetrics
and Gynecology,
Nara Medical University,
Kashihara-shi Nara 634, 'Habikino Hospital Osaka, 583, and 3 Third Department of Internal Medicine, National Defense Medical College, Saitama, 359, Japan
Received July 3, 1991
Humanmacrophage colony-stimulating factor (hM-CSF) concentrationdependently enhanced the secretion of human chorionic gonadotropin (hCG) by primary cultured human cytotrophoblastic cells and a human placental cell line, 3A-SubE (tPA30-1). Since this effect appeared 12 hours after the addition of hM-CSF and disappeared when protein synthesis was inhibited, it was surmised that hCG synthesis was enhanced by hM-CSF. When anti fms (hM-CSF receptor) antibody was added, hCG secretion by cultured human cytotrophoblasts in early pregnancy markedly decreased. These findings demonstrate that hM-CSF acts on the chorionic villous cells and promotes hCG synthesis by these cells. 0 1991Academic Pre**, Inc.
It
is already
known that
an M-CSF (CSF-1) receptor, the c-fms product receptor
(3),
physiological details
it action
the protooncogene c-fins,
is expressed in the placenta
is a single
that
M-CSF exerts
affinity
some type
of
chorionic villous cells, although the Recently, it has been shown that the content
of M-CSF in the pregnant murine uterus other tissues
in the endometrium are linked
is
(approximately
the expression of c-fms in the placenta is involved
Since
on the
remain obscure.
compared with
(1,2).
molecule and forms a high
thought
is
which encodes
(6,7),
in the differentiation
+ To whomcorrespondence should
be
remarkably lOOO-fold)
increased (4,5)
as
and that
and expression of II-CSF mRNA
leading to speculation or proliferation
that M-CSF
of trophoblastic
addressed. 0006-291X'91$1.50 1099
Copyright 0 1991 by Academic Press, Inc. All rights of reproduction in any form reserved.
Vol.
178,
No.
cells.
BIOCHEMICAL
3, 1991
In the present
by the chorionic
MATERIALS
HETBODS
AND
BIOPHYSICAL
we investigated
study,
hCG secretion
AND
villous
RESEARCH
whether
COMMUNICATIONS
M-CSF increases
cells.
1) ISOLATION AND CULTDRR OF CYTGTROPHOBLASTIC CRLLS Cytotrophoblastic cells were isolated by to the method of Rlinvnan et al.(s). After adjusting the obtained cells to a concentration of 2 x lO'/mL in 10% fetal calf serum supplemented with RPM1 1640 medium, the mixture was divided into 100 PII lots and poured into a 96-well flat plate which had been coated with laminin (Iwaki, Tokyo Japan). were cultured overnight at 37OC in a 5% CD, Then, the cells atmosphere. The next day, the supernatant was aspirated, and the cells cultured in serum-free HITES medium (RPM1 1640, 10 nM hydrocortisone, 5 ug/mk insulin, 100 ng transferrin, 30 nM sodium selenite). hM-CSF and anti fms antibody were added to the serum-free culture medium at the time of culture. In a flow cytometical analysis using Leu M3, the monocyte contamination rate was 65%. 2) hM-CSF, AND ANTI FNS ANTIBODY hM-CSF was obtained from Morinaga Milk Industry (Tokyo, Japan) in the form of a purified product obtained from human urine 9). The specific activity of this hM-CSF was l-2 x lO"U/mg as determined by the soft agar colony formation method using mouse bone marrow cells. Rabbit polyclonal antibody to fms was purchased from Cambridge Research Biochemicals (UK) and was added to the culture system to give a concentration of 1.0 ngIgG/mE. 3) CULTURE OF 3A-SubE (tPA30-1) CRLLS 3A-SubE (tPA30-1) cells are SV40 tsA30 virus transformed cell line that produces hCG only when cultured at 40°C (10). In the present experiment, the cells were cultured for 4 days at 40°C in RPM1 1640 medium supplemented with 10% FCS, and then cultured for an additional 2 days after changing the medium and adding hM-CSF. 4) DETRRMINATION OF hCG hCG was measured with an RLISA kit (Mochida Pharmaceuticals, Tokyo, Japan) using monoclonal antibodies against hCG a chain and 8 chain complex (11).
RESULTS Twenty-four hours human cytotrophoblast amount of
after adding hM-CSF, the amount of hCG in the culture supernatant increased with increasing
added hM-CSF,
2-100
ng/mll.
In the case of 3A-SubE the amount of hCG in the culture supernatant
(tPA30-1)
cells
increased Observing
depending on the concentration of added hM-CSF (Table the time course of the amount of hCG in the supernatant
the
as well,
from
human cytotrophoblast
culture
in
the
presence
of
1). of
20 rig/m!?, of
hM-CSF, a larger amount of hCG was found after 12 hours as compared with the supernatant of the control culture without hM-CSF (Fig 1). 1100
Vol.
178,
No.
3, 1991
BIOCHEMICAL
Table 1.
AND
mIU/mt 2190 2400 4230 5420
1) Mean f SD in triplicate
that
after
hM-CSF did
the addition of
via protein secretion
of
not
was
not
of
cytotrophoblasts
1021) 151 230 515
enhanced
cell by the
in
all
anti-fms
that
was examined
f f f f
on (Table
amount of within
0.2') 0.7 0.9 1.3
addition
hCG in
the initial
the induction
of hM-CSF is
6 h
when hM-CSF was
mediated that
hCG
added to
2).
of h&I-CSF on hCG secretion,
the
the secretion of hCG from 3). By adding hH-CSF (20 rig/me),
Fig. L Time course of the amount of hCG in the human cytotrophoblast culture supernatant in the presence of hM-CSF. 1101
the
of the hCG
by the observation
(Table
the influence antibody
the
culture
4 cases
cytotrophoblasts clarify
h
6.5 14.2 22.1 35.3
increase
This was supported
cycloheximide-treated To further
hCG/Z x lo4 cells/24
hM-CSF suggested
cytotrophoblasts synthesis.
tpA30-1
assay,
of the cytotrophoblast
synthesis
effect
f f f f
COMMUNICATIONS
trophoblasts
Human trophoblast
0 2 20 100
The finding
RESEARCH
The hCG concentration of primary cultured and tPA30-1 in the presence of M-CSF
Concentration of hI4-CSF (ng/m!Z)
supernatant
BIOPHYSICAL
Vol.
178,
No.
BIOCHEMICAL
3, 1991
Table
2.
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
Effects of hk!-CSF and cycloheximide on hCG production primary cultured human trophoblastic cells Medium
hkl-CSF1)
1 2 3 4
1910 810 540 590
Cycloheximide2)
by
cycloheximid+hH-CSF3)
hCG/2x 10' cells/24 h
mIU/mk
Case Case Case Case
AND
2940 1510 925 670
140 215 490 555
560 220 510 550
1) NI-CSF (20 ng/mL) was added to humantroPhoblastic cells for 24 h. 2) Cycloheximide (50 us/me) was added to human trophoblastic cells for 24 h. 3) hH-CSF (20 ng/mt) and cycloheximide (50 pg/mt) were added to humen trophoblastic cells for 24 h.
the
production
of
50% as compared
with
In contrast,
fms amb0ay effect
hCG by
cells
increased
to
182
f
no addition.
hCG synthesis
was added,
on the
cytotrophoblastic
although
was suppressed 53.9 f 21.6% when antithe
addition
rabbit
of
serum had no
of hCG secretion.
amount
DISCDSSION The
present
study
demonstrated
that
hM-CSF increases
the
synthesis
and secretion of hCG from the trophoblasts. The fact #at the same phenomenonwas also confirmed in the 3A-subE (tPA30-1) cells suggested that hM-CSF acts not only on monocytes, which are present in a very
small number, but also on trophoblasts.
Table
It
has been reported
3. The hCG levels in the supernatant of primary cultured trophoblastic cells with hM-CSF or anti-fms antibody
Medium
hBl-CSF (20 rig/me) mIU
135 282 2190 636 1910 800 540 430
anti fms Ab (1 w/me)
hCG/Z x lo* trophoblast 2214 495 4230 840 2940 1510 925 590
365 79 1824 397 609 n.t.a) 175 n.t.
a) not tested.
1102
Normal rabbit serum (1% volume) cells/24
h 130 210 2338 654 1875 830 n.t. n.t.
Vol.
178,
No.
that
3, 1991
M-CSF
However,
BIOCHEMICAL
is
a
since
AND
trophoblast
no increase
BIOPHYSICAL
RESEARCH
proliferation
factor
in DNA synthesis
rig/m% of hM-CSF was added to 3A-subE
COMMUNICATIONS
was
(tPA30-1)
in
found
cells
mice
12).
even when 20
(data
not shown),
the hM-CSF-induced enhancement of hCG secretion from the trophoblastic cells is probably not due to an increase in the number of trophoblasts. The production A-kinase play
mediated
some role
activates
enhanced
(15-17). as well,
receptor
and c-fms
may influence
(26).
by
known to
that
is thought
growth
tyrosine-specific that
C also
C may be activated,
and interestingly
cytokines,
hM-CSF that
in
resulting
factor
(EGF)
also
enough, both the EGF protein
kinase
tyrosine-specific
interleukin to promote
IL-l,
IL-6,
These cytokines and release
M-CSF are present It
and so it
Epidermal
suggesting
induced
has been reported
kinase
(19,20);
(23-25).
production
it
protein
have been reported
respectively linked
kinase
protein
mainly
protein
in
their kinase
hCG secretion.
Among other (IL-6)
with C (18),
possess
(21,22),
is
kinase
hCG production.
hCG secretion
trophoblasts
Recently,
protein
enhances cytoplasm
hCG from
by CAMP (13,14),
monocyte
the trophoblast in
of
involved
in
the
chorionic
villous
of hCG. that
regulation
the secretion
Also,
these of
aa
interleukin
and release
and hM-CSF secretion
probably
in the decidua
seems likely
1 (IL-l)
act it
in concert
IL-l,
closely IL-6
and
(5,27,28).
in the decidua
hCG secretion
of hCG,
to enhance the
is known that
and endometrium cytokines
are
6
and
may also
release
by
be the
cells.
REFERENCES 1)
Miller R., Shnon D.J., Adamson E.D., Tremblay J.M., Miiller D., Cline M-J., Verma I.M. (1983) Mol. Cell. Biol. 3: 1062-1069. 2) Hoshina M., Nishio A., Bo M., Biome I., Mochizuki M. (1985) Acta Obstet. Gynec. Jpn. 37: 2791-2798. 3) Guilbert L.J., Stanley E.R. (1986) J. Biol. Chem. 261: 4024-4032. 4) Bartocci A, Pollard J.W., Stanley R. (1986) J. Fxp. Med. 164: 956-961. 5) Pollard J.W., Bartocci A., Areci R., Orlofsky A., Landner M.B., Stanley R. (1987) Nature 330: 484-486. 6) Arceci R-J., Shanahan F., Stanley R., Polard J.W. (1989) Proc. Natl. Acad. Sci, USA. 86: 8818-8822. 7) Regenstreif L.J., Rossant J. (1989) Dev. Biol. 133: 284-294. 8) Kliman H.J., Nestler J.E., Sermasi E., Sanger J-M., Stranss J.F. (1986) Endocrinology. 118: 1567-1582. 9) Hanamura T.K., Motoyoshi K., Yoshida K., Saito M., Miura Y., Kawashima T., Nishide M., Takaku F. (1988) Blood. 72: 886-892. 10) Chou J.Y. (1978) Proc. Natl. Acad. Sci. USA. 75: 1409-1413. 11) Iwashita M., Watanabe M. Adachi T., Ohira A., Shinozaki Y., Takeda Y *, Sakamoto S. (1989) Placenta. 10: 103-112. 1103
Vol.
178,
No.
3, 1991
BIOCHEMICAL
AND
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
12) Athsnassakis I. Bleackey R.C., Peatkan V., Guilbert L., Barr P.J., Wagmann T.G. (1987) J. -01. 138: 37-44. 13) Chan J.Y., Wang S.S., Robinson J.C. (1978) J. Clin. Endocrinol. Metab. 47: 46-51. 14) Milsted A., Cox. R.P. Nilson H.J. (1987) DNA 6: 213-219. 15) Ritvos O., Jalkanen J., Huthaniemi I, Stenman U.H., Alfthan H., Ranta T. (1987) Endocrinology 120: 1521-1526. 16) Andersen B., Milsted A, Kennedy G., Nilson J.H. (1988) J. Biol. Chem. 263: 15578-15583. 17) Ritvos O., Butzow R., Jalkanen J., Stenman U.H., Huhtaniemi I., Ranta T. (1988) Mel Cell Endocrinol. 56: 165-169. 18) Imamura K., Dianoux A., Nskamura T., Kufe D. (1990) EMBC J, 9: 2423- 2429. 19) Lai W.H., Guyda H.J. (1984) J. Clin. Endocrinol. Hetab. 58: 344-352. 20) Ritvos O., Jalkanen J., Pekonen F., Stenman U.H., Ranta. T. (1988) Endocrinology 123: 859-865. 21) Ullrich A., Coussens L., Hayflick J-S., Dull T.J., Gray A., Tam A-W., Lee J., Yaden Y., Liberman T.A., Schlessinger J., Downward J Mayes E.L.V., Whittle N., Waterfield, P.H., Seeburg P.H. (1684) Nature 309: 418-425. 22) Sherr C.J., Rettenmier C-W., Sacca R., Roussel M.F., Look A.T., Stanley E.R. (1985) Cell 41: 665-676. 23) Yagel S., Lala P., Powell W.A., Casper R.F. (1989) J. Clin. Endocrinol. Metab. 68: 992-995. 24) Silen M.L., Firpo A., Francus T., Klein R.F., Lowry S.F. (1989) Biochem. Biophys. Res. Connrmn. 164: 284-289. 25) Nishino E., Matsuzaki N., Masuhiro K., Kameda T., Taniguchi T., Takagi T., Saji F., Tanizawa 0. J. Clin. Endocrinol ,. Metab. (1990) 71: 436-441. 26) Haworth C. (1989) Blood Rev. 3: 263-268. 27) Remero R., Wu Y.K., Brody D.T., Oyarzun E., Duff G.W'., Durum S.K. (1989) Obstet. Gynecol. 73: 31-34. 28) Romero R., Avila C., Santhanam U., Sehgel P.B. (1990 ) J. Clin. Invest. 85: 1392-1400.
1104
178,
No. 15,
3, 1991
BIOCHEMICAL
BIOPHYSICAL
AND
RESEARCH
COMMUNICATIONS
Pages 1099-1104
1991
ENHANCING EFFECTSOF HUMANMACROPHAGE COLONY-STIMULATING FACTOR ON THE SECRRTION OF HUMANCHORIONICGONADOTROPIN BY HUMANCHORIONICVILLOUS CJILLSAND tPA30-1 CKLLS
Shigeru
1,+
Saito
, Hami Saito' , Kazuo Motoyoshi3 , and Motohiko Ichijo'
IDepartment of Obstetrics
and Gynecology,
Nara Medical University,
Kashihara-shi Nara 634, 'Habikino Hospital Osaka, 583, and 3 Third Department of Internal Medicine, National Defense Medical College, Saitama, 359, Japan
Received July 3, 1991
Humanmacrophage colony-stimulating factor (hM-CSF) concentrationdependently enhanced the secretion of human chorionic gonadotropin (hCG) by primary cultured human cytotrophoblastic cells and a human placental cell line, 3A-SubE (tPA30-1). Since this effect appeared 12 hours after the addition of hM-CSF and disappeared when protein synthesis was inhibited, it was surmised that hCG synthesis was enhanced by hM-CSF. When anti fms (hM-CSF receptor) antibody was added, hCG secretion by cultured human cytotrophoblasts in early pregnancy markedly decreased. These findings demonstrate that hM-CSF acts on the chorionic villous cells and promotes hCG synthesis by these cells. 0 1991Academic Pre**, Inc.
It
is already
known that
an M-CSF (CSF-1) receptor, the c-fms product receptor
(3),
physiological details
it action
the protooncogene c-fins,
is expressed in the placenta
is a single
that
M-CSF exerts
affinity
some type
of
chorionic villous cells, although the Recently, it has been shown that the content
of M-CSF in the pregnant murine uterus other tissues
in the endometrium are linked
is
(approximately
the expression of c-fms in the placenta is involved
Since
on the
remain obscure.
compared with
(1,2).
molecule and forms a high
thought
is
which encodes
(6,7),
in the differentiation
+ To whomcorrespondence should
be
remarkably lOOO-fold)
increased (4,5)
as
and that
and expression of II-CSF mRNA
leading to speculation or proliferation
that M-CSF
of trophoblastic
addressed. 0006-291X'91$1.50 1099
Copyright 0 1991 by Academic Press, Inc. All rights of reproduction in any form reserved.
Vol.
178,
No.
cells.
BIOCHEMICAL
3, 1991
In the present
by the chorionic
MATERIALS
HETBODS
AND
BIOPHYSICAL
we investigated
study,
hCG secretion
AND
villous
RESEARCH
whether
COMMUNICATIONS
M-CSF increases
cells.
1) ISOLATION AND CULTDRR OF CYTGTROPHOBLASTIC CRLLS Cytotrophoblastic cells were isolated by to the method of Rlinvnan et al.(s). After adjusting the obtained cells to a concentration of 2 x lO'/mL in 10% fetal calf serum supplemented with RPM1 1640 medium, the mixture was divided into 100 PII lots and poured into a 96-well flat plate which had been coated with laminin (Iwaki, Tokyo Japan). were cultured overnight at 37OC in a 5% CD, Then, the cells atmosphere. The next day, the supernatant was aspirated, and the cells cultured in serum-free HITES medium (RPM1 1640, 10 nM hydrocortisone, 5 ug/mk insulin, 100 ng transferrin, 30 nM sodium selenite). hM-CSF and anti fms antibody were added to the serum-free culture medium at the time of culture. In a flow cytometical analysis using Leu M3, the monocyte contamination rate was 65%. 2) hM-CSF, AND ANTI FNS ANTIBODY hM-CSF was obtained from Morinaga Milk Industry (Tokyo, Japan) in the form of a purified product obtained from human urine 9). The specific activity of this hM-CSF was l-2 x lO"U/mg as determined by the soft agar colony formation method using mouse bone marrow cells. Rabbit polyclonal antibody to fms was purchased from Cambridge Research Biochemicals (UK) and was added to the culture system to give a concentration of 1.0 ngIgG/mE. 3) CULTURE OF 3A-SubE (tPA30-1) CRLLS 3A-SubE (tPA30-1) cells are SV40 tsA30 virus transformed cell line that produces hCG only when cultured at 40°C (10). In the present experiment, the cells were cultured for 4 days at 40°C in RPM1 1640 medium supplemented with 10% FCS, and then cultured for an additional 2 days after changing the medium and adding hM-CSF. 4) DETRRMINATION OF hCG hCG was measured with an RLISA kit (Mochida Pharmaceuticals, Tokyo, Japan) using monoclonal antibodies against hCG a chain and 8 chain complex (11).
RESULTS Twenty-four hours human cytotrophoblast amount of
after adding hM-CSF, the amount of hCG in the culture supernatant increased with increasing
added hM-CSF,
2-100
ng/mll.
In the case of 3A-SubE the amount of hCG in the culture supernatant
(tPA30-1)
cells
increased Observing
depending on the concentration of added hM-CSF (Table the time course of the amount of hCG in the supernatant
the
as well,
from
human cytotrophoblast
culture
in
the
presence
of
1). of
20 rig/m!?, of
hM-CSF, a larger amount of hCG was found after 12 hours as compared with the supernatant of the control culture without hM-CSF (Fig 1). 1100
Vol.
178,
No.
3, 1991
BIOCHEMICAL
Table 1.
AND
mIU/mt 2190 2400 4230 5420
1) Mean f SD in triplicate
that
after
hM-CSF did
the addition of
via protein secretion
of
not
was
not
of
cytotrophoblasts
1021) 151 230 515
enhanced
cell by the
in
all
anti-fms
that
was examined
f f f f
on (Table
amount of within
0.2') 0.7 0.9 1.3
addition
hCG in
the initial
the induction
of hM-CSF is
6 h
when hM-CSF was
mediated that
hCG
added to
2).
of h&I-CSF on hCG secretion,
the
the secretion of hCG from 3). By adding hH-CSF (20 rig/me),
Fig. L Time course of the amount of hCG in the human cytotrophoblast culture supernatant in the presence of hM-CSF. 1101
the
of the hCG
by the observation
(Table
the influence antibody
the
culture
4 cases
cytotrophoblasts clarify
h
6.5 14.2 22.1 35.3
increase
This was supported
cycloheximide-treated To further
hCG/Z x lo4 cells/24
hM-CSF suggested
cytotrophoblasts synthesis.
tpA30-1
assay,
of the cytotrophoblast
synthesis
effect
f f f f
COMMUNICATIONS
trophoblasts
Human trophoblast
0 2 20 100
The finding
RESEARCH
The hCG concentration of primary cultured and tPA30-1 in the presence of M-CSF
Concentration of hI4-CSF (ng/m!Z)
supernatant
BIOPHYSICAL
Vol.
178,
No.
BIOCHEMICAL
3, 1991
Table
2.
BIOPHYSICAL
RESEARCH
COMMUNICATIONS
Effects of hk!-CSF and cycloheximide on hCG production primary cultured human trophoblastic cells Medium
hkl-CSF1)
1 2 3 4
1910 810 540 590
Cycloheximide2)
by
cycloheximid+hH-CSF3)
hCG/2x 10' cells/24 h
mIU/mk
Case Case Case Case
AND
2940 1510 925 670
140 215 490 555
560 220 510 550
1) NI-CSF (20 ng/mL) was added to humantroPhoblastic cells for 24 h. 2) Cycloheximide (50 us/me) was added to human trophoblastic cells for 24 h. 3) hH-CSF (20 ng/mt) and cycloheximide (50 pg/mt) were added to humen trophoblastic cells for 24 h.
the
production
of
50% as compared
with
In contrast,
fms amb0ay effect
hCG by
cells
increased
to
182
f
no addition.
hCG synthesis
was added,
on the
cytotrophoblastic
although
was suppressed 53.9 f 21.6% when antithe
addition
rabbit
of
serum had no
of hCG secretion.
amount
DISCDSSION The
present
study
demonstrated
that
hM-CSF increases
the
synthesis
and secretion of hCG from the trophoblasts. The fact #at the same phenomenonwas also confirmed in the 3A-subE (tPA30-1) cells suggested that hM-CSF acts not only on monocytes, which are present in a very
small number, but also on trophoblasts.
Table
It
has been reported
3. The hCG levels in the supernatant of primary cultured trophoblastic cells with hM-CSF or anti-fms antibody
Medium
hBl-CSF (20 rig/me) mIU
135 282 2190 636 1910 800 540 430
anti fms Ab (1 w/me)
hCG/Z x lo* trophoblast 2214 495 4230 840 2940 1510 925 590
365 79 1824 397 609 n.t.a) 175 n.t.
a) not tested.
1102
Normal rabbit serum (1% volume) cells/24
h 130 210 2338 654 1875 830 n.t. n.t.
Vol.
178,
No.
that
3, 1991
M-CSF
However,
BIOCHEMICAL
is
a
since
AND
trophoblast
no increase
BIOPHYSICAL
RESEARCH
proliferation
factor
in DNA synthesis
rig/m% of hM-CSF was added to 3A-subE
COMMUNICATIONS
was
(tPA30-1)
in
found
cells
mice
12).
even when 20
(data
not shown),
the hM-CSF-induced enhancement of hCG secretion from the trophoblastic cells is probably not due to an increase in the number of trophoblasts. The production A-kinase play
mediated
some role
activates
enhanced
(15-17). as well,
receptor
and c-fms
may influence
(26).
by
known to
that
is thought
growth
tyrosine-specific that
C also
C may be activated,
and interestingly
cytokines,
hM-CSF that
in
resulting
factor
(EGF)
also
enough, both the EGF protein
kinase
tyrosine-specific
interleukin to promote
IL-l,
IL-6,
These cytokines and release
M-CSF are present It
and so it
Epidermal
suggesting
induced
has been reported
kinase
(19,20);
(23-25).
production
it
protein
have been reported
respectively linked
kinase
protein
mainly
protein
in
their kinase
hCG secretion.
Among other (IL-6)
with C (18),
possess
(21,22),
is
kinase
hCG production.
hCG secretion
trophoblasts
Recently,
protein
enhances cytoplasm
hCG from
by CAMP (13,14),
monocyte
the trophoblast in
of
involved
in
the
chorionic
villous
of hCG. that
regulation
the secretion
Also,
these of
aa
interleukin
and release
and hM-CSF secretion
probably
in the decidua
seems likely
1 (IL-l)
act it
in concert
IL-l,
closely IL-6
and
(5,27,28).
in the decidua
hCG secretion
of hCG,
to enhance the
is known that
and endometrium cytokines
are
6
and
may also
release
by
be the
cells.
REFERENCES 1)
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