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Radioactive estradiol accumulation in endometriosis of the rhesus monkey
Radioactive estradiol accumulation endometriosis A.
J.
W.
U.
G.
VAN
New
in
of the rhesus monkey
EISENFELD
Haven,
GARDNER WAGENEN Connecticut
$H-e&radio1 was administered
intravenously to 7 rhesus monkeys and the concentration of radioactivity in tissues measured one or two hours later. Four of the monkeys had endometriosis; in three the disease followed pelvic irradiation several years previously. High concentrations of radioactivity were found in the endometrium, myometrium, endometriosis, and adenomyosis relative to the concentrations in plasma, fat, or muscle.
ENDOMETRIOSIS of the rhesus monkey occurs spontaneously1 and in high incidence following uterotomy2-4 or pelvic irradiation.6 To date 8 of 13 monkeys that received 600 to 900 roentgen units of x-ray through a 6 by 8 cm. port to the pelvis have developed endometriosis.6 Endometriosis of the monkey resembles the disease in women both in its gross appearance and its histopathology. It is unknown whether endometriosis develops by peritoneal metaplasia or following implantation of endometrial tissue in aberrant sites.7 In women the disease is unknown before menarche and is uncommon following menopause.8 Experimentally an extension or invasion of endometrium into adjacent tissues has been described when endometrium has been fixed in juxtaposition to other tissues99 lo Endometrial explants in castrated monkeys increase in size and exhibit glandular development with estrogen From the Departments and Internal Medicine, Obstetrics-Gynecology, School of Medicine.
or estrogen plus intermittent progesterone therapy.ll To provide evidence consistent with a direct effect of estrogens on endometriosis we have studied the accumulation of estradiol in endometriosis of the monkey after intravenous administration of 3H-estradiol. It is known that high concentrations of 3H-estradiol accumulate in the rat’s uterus.12
Materials and methods Radioactive 17P-estradiol (6, 7-3H, 48 C./mmole, New England Nuclear) one pg per kilogram of body weight in 1 ml. of 50 per cent ethanol, was injected into a saphenous vein during a one-minute period. The animals were anesthetized with Nembutal shortly before injection and remained anesthetized in a warm room until killed by exsanguination 1 or 2 hours later. Tissues were removed rapidly, placed on ice, weighed, and frozen until analyzed. As described previously,13 the tissues were homogenized in 2 ml. of chilled water and extracted with 10 ml. of 95% toluene: 5 per cent isoamyl alcohol. Unconjugated estrogens were extracted into the organic phase. After shaking, 5 ml. of the organic layer was added to 10 ml. of PPO-POPOP in toluene for liquid scintillation radioactive counting (Packard 3320 ef-
of Pharmacology Anatomy, and Yale Unrversity
Supported in part by Grants Ca 00429 and Ca 00343 from the National Cancer Institute and HD 02498 from the Child Health and Development Institute, United States Public Health Service, and by the Ford Foundation. 124
Volume Number
109 1
Radioactive
ficiency 45 per cent). Two or more samples were assayed from each tissue. Results were expressed as ppcuries of radioactivity per milligram of wet tissue weight corrected for the portion counted and extraction efficiency (80 per cent). The radioactivity in the organic extract from the same tissues from several animals was pooled and characterized in two chromatographic systems: the Bush 3 system and the system with formamide im-
Table I. Reproductive No.
of animals
1 endometriosis
history
of the monkeys
Age at irradiation
-
Age
estradiol
in endometriosis
125
pregnated paper with o-dichlorobenzene as mobile phase.14 The reproductive histories of the monkeys given SH-estradiol are summarized in Table I. The first monkey with endometriosis (No. 1) had not received irradiation. She was pregnant in 1957 and 1958 with fetuses removed by uterostomy on days 46, 150, and 100. In 1960 gonadotropins were given and
given 3H-estradiol
at death (yr.)
ca. 15
Summary reproductive
of history
5 pregnancies (3 early cesare-
Body
weight (Kg.)
7.6
ans) 2 normal
3 4 5 6 7
normal normal endometriosis endometriosis endometriosis
-
15 months 24 months 24 months
17% 5% ca. l&i ca.
21 21
6 pregnancies 1 pregnancy (induced abortion) No pregnancies known 2 pregnancies, both aborted 2 pregnancies, both aborted 1 pregnancy aborted
6.7 6.4 4.2 10.7 9.4 9.7
Fig. 1. Endometriosis in a nonirradiated monkey. Area of endometrium-like epithelium and stroma overlying a fibrous surface in the rectosigmoid area near the point of endometrial junction with the large bowel epithelium. (x310.)
126
Eisenfeld,
Gardner,
and
van
Wagenen
Amer.
one ovary was removed. Another pregnancy was completed in 1962. Several different experiments with contraceptive substances were done during 1964-1966. For the last several months the monkey had irregular cycles and both anal and vaginal bleeding. At necropsy in 1967 the entire uterine area was covered by adhesions of omentum and viscera to the upper surface of the bladder and adjacent body wall. Over the uterus and surrounding its free dorsal and cranial surface was a cyst containing viscous brownish material and measuring approximately 25 mm. in width. It contained polypoid masses measuring 22 by 13 by 15 mm. and 5 by 9 mm. The cyst was continuous with a fistula through the wall of the rectosigmoid junction. The cystic extrauterine structures were lined by endometrium-like tissue with epithelial cells varying from tall columnar to cuboidal arranged in glandular or papillary structures, the latter projecting into cysts. The stroma ranged from endometrium-like
Fig. 2. Endometriosis in a fibrous stroma. (x310.)
an irradiated
monkey
January J. Obstet.
1, 1971 Gynec.
(Fig. 1)) in places even containing coiled arteries, to largely fibrous. Glandular endometrial tissue also invaded the adipose tissue of the omentum, the serosa and muscle of the bladder, and lined a portion of the fistula into the rectosigmoid. The endometrium appeared typical for about the twenty-second day of the menstrual cycle. The myometrium contained endometrial glands and some endometrium-like stroma (adenomyosis) . Some areas of the endometriosis and adenomyosis had numerous erythrocytes in the interstitium and within a few of the glandlike structures. The next monkey with endometriosis (No. 5) received 600 roentgens to the pelvis at 1.5 months of age, about 17 years before the present experiment. This monkey spontaneously aborted twice. She had irregular menstrual cycles for the last 8 years. At necropsy the greater omentum was attached over the upper surface of the bladder. When detached, cysts containing viscous chocolate-
showing
epithelium-lined
tubules
surrounded
by
v01u!me Number
Radioaciive
109 1
colored fluid were exposed. The cystic struttures were lined by endometrium-like tissue in some places with endometrium-like stroma, in other places abutting directly against fibrocollagenous (Fig. 2) or adipose tissue. The bladder wall was invaded by the endometriosis.A tumor mass,25 by 20 by 24 mm., apparently completely separated from the cervix, was in the position of the uterus. The mass was composed of endometriumlike epithelium and stroma with remnants of myometrial tissue (adenomyosis) (Fig. 3). No normal uterine tissue was found. Monkey No. 6 was two years old when irradiated with 600 r and was killed 19 years later. During the postirradiation period she had two pregnancies that terminated in abortion. During the last two years she had almost continuous vaginal bleeding following 21/z years of amenorrhea. In the few months prior to autopsy an enlarging mass was palpable in the lower abdomen. At necropsy cysts and polyps lined by endomet&m-like tissue were adherent to the omentum and bladder. No normal uterine tissuewas found. In the position of the uterus were two large massesthat were histologically diagnosed as adenomyosis. A discrete 12 by 13 mm. nodule was found on the mesentery of the left sigmoid colon. Histologically this was composedof tall columnar epithelium in
Fig. 3. The uterine-like like
tissue
with
small
ma33
areas
of an irradiated of smooth muscle
estradiol
in endometriosis
127
endometrium-like and fibrous stroma (endometrioma) (Fig. 4). Monkey No. 7 was approximately 24 months of age when irradiated (600 r) . She had prolonged periods of vaginal bleeding for the last 2 years, a palpable abdominal mass,and a frozen pelvis. At necropsy at 21 years of age the animal had endometriosis with chocolate cysts, adenomyosis (Fig. 5), and a fibroma. No normal uterine tissuewas seenin the sectionsexamined. Three monkeys without endometriosis were studied. Monkeys Nos. 2 and 3 were used in contraceptive drug trials ending 3 years and 1.5 years before the present experiment. Monkey No. 4 was in the colony for only a few months. These monkeys without endometriosishad regular menstrual cycles. Histologically the endometriosis and adenomyosis in the non- and irradiated monkeys had glands arranged in a disorganized fashion but the cells were not anaplastic. Although the endometriosis was locally invasive, distant metastaseswere not found. None of the irradiated monkeys was used in other experiments. Results
After intravenous administration of 3Hestradiol, high levels of radioactivity were found in endometriosis and adenomyosis
monkey consisted predominantly (adenomyosis) . (x5.)
of endometrium-
128
Eisenfeld,
Gardner,
and
van
Wagenen
Fig. 4. A section from the endometrioma on the side opposite the epithelial surface.
Amer
attached (x105.)
Fig. 5. Section through adenomyosis. Myometrial some places with some endometrial stroma, in other (x105.)
to the mesentery.
A coiled
artery
January J. Obstet.
is located
areas were invaded by uterine glands places in direct contact with myometrium.
in
1, 1971 Gynec.
Volume 109 Number 1
Radioactive
Table II. Radioactivity
in tissues of the monkey after intravenous Monkey 1, uterostomy
Day of menstrual cycle Hours after injection Endometrium Myometrium Endometriosis Adenomyosis Endometrioma Skeletal muscle Subcutaneous fat Plasma
estradiol
Monkey
Monkey 3, normal
2,
normal 1
1,103
3 1 1,014 603
Monkey 4, normal
14 28:
73;
242
291
Monkey 6, irradiated
2
15 if
;:
58 30 52
27
2
Comment
The present data indicate the high accumulation of radioactive estradiol in endometrial tissue in aberrant sites. This may be due to the presence of binding macromole-
Ave. at 2 hr.
2
394 394 417 20 51 36
Monkeys were injected with 1 fig of *H-estradiol per kilogram intravenously and radioactivity one or two hours later. Radioactivity is expressed as pFurie per milligram of wet tissue weight.
(Table II). The levels averaged as high as in the normal endometrium. All the radioactivity in endometriosis, adenomyosis, endometriomas, endometrium, and myometrium and 80 per cent from plasma co-chromatographed with authentic estradiol. The radioactivity in endometriosis, adenomyosis, endometrium, and myometrium is significantly higher (p < 0.05) than the radioactivity in skeletal muscle, subcutaneous fat, or plasma. The concentration gradients relative to plasma were endometriosis 11, adenomyosis 17, endometrium 16, and myometrium 8. The concentration of estradiol in endometriosis and adenomyosis was considerably higher (at least threefold) than the radioactive concentration in a wide variety of extrauterine tissues.15 In general, the radioactivity was higher in portions of endometriosis that were rich in glandular tissue and lower in portions containing substantial amounts of fibrous or fatty tissue.
Monkey 7, irradiated
9
652 639 77 33 120
129
SH-estradiol
Monkey 5, irradiated
543 908 64 57 61
in endometriosis
258 631 20 49 22 in
514 267 435 555 27 48 33
tissues was measured
cules with a high affinity for estradiol in endometriosis. In uteri of rats the accumulation in vivo is limited in capacity,16 and has a high specificity for drugs with estrogenic and antiestrogenic activity.l?> I8 This accumulation appears to be due to protein-containing macromolecules which tightly bind estradioLl@The first interaction of estradiol with the uterine protein appears to occur in the cytoplasm with the possiblesubsequenttransfer of the estrogen and cytoplasmic protein into the nucleus20 where it attaches to chromatin.21 The estrogen-protein macromolecule complex is thought to regulate gene activity promoting RNA synthesiswith subsequentincreasesin protein synthesis,tissuegrowth, and cellular division.22 The present finding of high accumulation of estradiol in endometriosis is compatible with the possibility that a similar sequence of biochemical events may be operative in endometriosis. We thank Mrs. R. S&none, Mr. Joseph Negri, and Miss Dorothy Butzko for their excellent
technical
assistance.
REFERENCES
1. Kliiver, Gynec. 2. Hartman, Gynec.
H., and Bartelmez, G. W.: Surg. Obstet. 92: 650, 1957. C. G.: Western J. Surg. Obstet. 52: 87, 1944.
3. 4. 5.
Pickering, McCann, OBSTET. Gardner,
D. E.: Folia primat. 8: 169, 1968. T. O., and Myers, R. E.: AMER. J. GYNEC. 106: 516, 1970. W. U., and van Wagenen, G.: IX
130
Eisenfeld, Gordner,
January 1, 1971 Amer. J. Obstet. Gynec.
and van Wogenen
International Cancer Congress, Tokyo, Abst. 0201, 1966. G., and Gardner, W. U.: In 6. van Wagenen, preparation. 7. Baird, Sir Dugald, Combined Textbook of Obstetrics and Gvnecoloav. Edinburgh. 1969, E. & S. Livingstone, Ltd., p. 821. 8. Kistner, R. W.: The Use of Progestins in Obstetrics and Gynecology, Chicago, 1969, Year Book Medical Publishers, Inc. 9. van Wagenen, G., and Morse, A. H.: Endocrinology 27: 268, 1940. 10. Te Linde, R. W., and Scott, R. B.: AMER. J. QBSTET. GYNEC. 60: 1147, 1950. 11. Scott, R. B., and Wharton, L. R.: AMER. J. OBSTET. GYNEC. 74: 852, 1957. 12. Jensen, E. V., and Jacobson, H. I.: Recent Progr. Hormone Res. 18: 387, 1962. 13. Eisenfeld, A. J., and Axelrod, J.: J. Pharmacol. Exp. Ther. 150: 469, 1965. -r,
-,
_
14. 15. 16. 17. 18. 19. 20.
21. 22.
Axelrod, L. R.: J. Biol. Chem. 201: 59, 1953. Eisenfeld, A. J., Gardner, W. U., and van Wagenen, G.: In preparation. Eisenfeld, A. J.: Biochim. Biophys. Acta 136: 498, 1967. Eisenfeld, A. J., and Axelrod, J.: Endocrinology 79: 38, 1966. Eisenfeld, A. J., and Axelrod, J.: Biochem. Pharmacol. 16: 1781, 1967, Toft, D., Shyamala, -G., and Gorski, J.: Proc. Nat. Acad. Sci., U.S.A. 57: 1740. 1967. Jensen, E. V.,. Suzuki, T., Kawashima, T., Stumpf, W., Jungblut, P., and DeSombre, E.: Proc. Nat. Acad- Sci., U.S.A. 50: 632, 1968. Maurer, H. R., and Chalklev. G. R.: YT. Molec. Biol. 27: 431, 1967. ” Hamilton, T. H.: Science 161: 649, 1968.
J.
W.
U.
G.
VAN
New
in
of the rhesus monkey
EISENFELD
Haven,
GARDNER WAGENEN Connecticut
$H-e&radio1 was administered
intravenously to 7 rhesus monkeys and the concentration of radioactivity in tissues measured one or two hours later. Four of the monkeys had endometriosis; in three the disease followed pelvic irradiation several years previously. High concentrations of radioactivity were found in the endometrium, myometrium, endometriosis, and adenomyosis relative to the concentrations in plasma, fat, or muscle.
ENDOMETRIOSIS of the rhesus monkey occurs spontaneously1 and in high incidence following uterotomy2-4 or pelvic irradiation.6 To date 8 of 13 monkeys that received 600 to 900 roentgen units of x-ray through a 6 by 8 cm. port to the pelvis have developed endometriosis.6 Endometriosis of the monkey resembles the disease in women both in its gross appearance and its histopathology. It is unknown whether endometriosis develops by peritoneal metaplasia or following implantation of endometrial tissue in aberrant sites.7 In women the disease is unknown before menarche and is uncommon following menopause.8 Experimentally an extension or invasion of endometrium into adjacent tissues has been described when endometrium has been fixed in juxtaposition to other tissues99 lo Endometrial explants in castrated monkeys increase in size and exhibit glandular development with estrogen From the Departments and Internal Medicine, Obstetrics-Gynecology, School of Medicine.
or estrogen plus intermittent progesterone therapy.ll To provide evidence consistent with a direct effect of estrogens on endometriosis we have studied the accumulation of estradiol in endometriosis of the monkey after intravenous administration of 3H-estradiol. It is known that high concentrations of 3H-estradiol accumulate in the rat’s uterus.12
Materials and methods Radioactive 17P-estradiol (6, 7-3H, 48 C./mmole, New England Nuclear) one pg per kilogram of body weight in 1 ml. of 50 per cent ethanol, was injected into a saphenous vein during a one-minute period. The animals were anesthetized with Nembutal shortly before injection and remained anesthetized in a warm room until killed by exsanguination 1 or 2 hours later. Tissues were removed rapidly, placed on ice, weighed, and frozen until analyzed. As described previously,13 the tissues were homogenized in 2 ml. of chilled water and extracted with 10 ml. of 95% toluene: 5 per cent isoamyl alcohol. Unconjugated estrogens were extracted into the organic phase. After shaking, 5 ml. of the organic layer was added to 10 ml. of PPO-POPOP in toluene for liquid scintillation radioactive counting (Packard 3320 ef-
of Pharmacology Anatomy, and Yale Unrversity
Supported in part by Grants Ca 00429 and Ca 00343 from the National Cancer Institute and HD 02498 from the Child Health and Development Institute, United States Public Health Service, and by the Ford Foundation. 124
Volume Number
109 1
Radioactive
ficiency 45 per cent). Two or more samples were assayed from each tissue. Results were expressed as ppcuries of radioactivity per milligram of wet tissue weight corrected for the portion counted and extraction efficiency (80 per cent). The radioactivity in the organic extract from the same tissues from several animals was pooled and characterized in two chromatographic systems: the Bush 3 system and the system with formamide im-
Table I. Reproductive No.
of animals
1 endometriosis
history
of the monkeys
Age at irradiation
-
Age
estradiol
in endometriosis
125
pregnated paper with o-dichlorobenzene as mobile phase.14 The reproductive histories of the monkeys given SH-estradiol are summarized in Table I. The first monkey with endometriosis (No. 1) had not received irradiation. She was pregnant in 1957 and 1958 with fetuses removed by uterostomy on days 46, 150, and 100. In 1960 gonadotropins were given and
given 3H-estradiol
at death (yr.)
ca. 15
Summary reproductive
of history
5 pregnancies (3 early cesare-
Body
weight (Kg.)
7.6
ans) 2 normal
3 4 5 6 7
normal normal endometriosis endometriosis endometriosis
-
15 months 24 months 24 months
17% 5% ca. l&i ca.
21 21
6 pregnancies 1 pregnancy (induced abortion) No pregnancies known 2 pregnancies, both aborted 2 pregnancies, both aborted 1 pregnancy aborted
6.7 6.4 4.2 10.7 9.4 9.7
Fig. 1. Endometriosis in a nonirradiated monkey. Area of endometrium-like epithelium and stroma overlying a fibrous surface in the rectosigmoid area near the point of endometrial junction with the large bowel epithelium. (x310.)
126
Eisenfeld,
Gardner,
and
van
Wagenen
Amer.
one ovary was removed. Another pregnancy was completed in 1962. Several different experiments with contraceptive substances were done during 1964-1966. For the last several months the monkey had irregular cycles and both anal and vaginal bleeding. At necropsy in 1967 the entire uterine area was covered by adhesions of omentum and viscera to the upper surface of the bladder and adjacent body wall. Over the uterus and surrounding its free dorsal and cranial surface was a cyst containing viscous brownish material and measuring approximately 25 mm. in width. It contained polypoid masses measuring 22 by 13 by 15 mm. and 5 by 9 mm. The cyst was continuous with a fistula through the wall of the rectosigmoid junction. The cystic extrauterine structures were lined by endometrium-like tissue with epithelial cells varying from tall columnar to cuboidal arranged in glandular or papillary structures, the latter projecting into cysts. The stroma ranged from endometrium-like
Fig. 2. Endometriosis in a fibrous stroma. (x310.)
an irradiated
monkey
January J. Obstet.
1, 1971 Gynec.
(Fig. 1)) in places even containing coiled arteries, to largely fibrous. Glandular endometrial tissue also invaded the adipose tissue of the omentum, the serosa and muscle of the bladder, and lined a portion of the fistula into the rectosigmoid. The endometrium appeared typical for about the twenty-second day of the menstrual cycle. The myometrium contained endometrial glands and some endometrium-like stroma (adenomyosis) . Some areas of the endometriosis and adenomyosis had numerous erythrocytes in the interstitium and within a few of the glandlike structures. The next monkey with endometriosis (No. 5) received 600 roentgens to the pelvis at 1.5 months of age, about 17 years before the present experiment. This monkey spontaneously aborted twice. She had irregular menstrual cycles for the last 8 years. At necropsy the greater omentum was attached over the upper surface of the bladder. When detached, cysts containing viscous chocolate-
showing
epithelium-lined
tubules
surrounded
by
v01u!me Number
Radioaciive
109 1
colored fluid were exposed. The cystic struttures were lined by endometrium-like tissue in some places with endometrium-like stroma, in other places abutting directly against fibrocollagenous (Fig. 2) or adipose tissue. The bladder wall was invaded by the endometriosis.A tumor mass,25 by 20 by 24 mm., apparently completely separated from the cervix, was in the position of the uterus. The mass was composed of endometriumlike epithelium and stroma with remnants of myometrial tissue (adenomyosis) (Fig. 3). No normal uterine tissue was found. Monkey No. 6 was two years old when irradiated with 600 r and was killed 19 years later. During the postirradiation period she had two pregnancies that terminated in abortion. During the last two years she had almost continuous vaginal bleeding following 21/z years of amenorrhea. In the few months prior to autopsy an enlarging mass was palpable in the lower abdomen. At necropsy cysts and polyps lined by endomet&m-like tissue were adherent to the omentum and bladder. No normal uterine tissuewas found. In the position of the uterus were two large massesthat were histologically diagnosed as adenomyosis. A discrete 12 by 13 mm. nodule was found on the mesentery of the left sigmoid colon. Histologically this was composedof tall columnar epithelium in
Fig. 3. The uterine-like like
tissue
with
small
ma33
areas
of an irradiated of smooth muscle
estradiol
in endometriosis
127
endometrium-like and fibrous stroma (endometrioma) (Fig. 4). Monkey No. 7 was approximately 24 months of age when irradiated (600 r) . She had prolonged periods of vaginal bleeding for the last 2 years, a palpable abdominal mass,and a frozen pelvis. At necropsy at 21 years of age the animal had endometriosis with chocolate cysts, adenomyosis (Fig. 5), and a fibroma. No normal uterine tissuewas seenin the sectionsexamined. Three monkeys without endometriosis were studied. Monkeys Nos. 2 and 3 were used in contraceptive drug trials ending 3 years and 1.5 years before the present experiment. Monkey No. 4 was in the colony for only a few months. These monkeys without endometriosishad regular menstrual cycles. Histologically the endometriosis and adenomyosis in the non- and irradiated monkeys had glands arranged in a disorganized fashion but the cells were not anaplastic. Although the endometriosis was locally invasive, distant metastaseswere not found. None of the irradiated monkeys was used in other experiments. Results
After intravenous administration of 3Hestradiol, high levels of radioactivity were found in endometriosis and adenomyosis
monkey consisted predominantly (adenomyosis) . (x5.)
of endometrium-
128
Eisenfeld,
Gardner,
and
van
Wagenen
Fig. 4. A section from the endometrioma on the side opposite the epithelial surface.
Amer
attached (x105.)
Fig. 5. Section through adenomyosis. Myometrial some places with some endometrial stroma, in other (x105.)
to the mesentery.
A coiled
artery
January J. Obstet.
is located
areas were invaded by uterine glands places in direct contact with myometrium.
in
1, 1971 Gynec.
Volume 109 Number 1
Radioactive
Table II. Radioactivity
in tissues of the monkey after intravenous Monkey 1, uterostomy
Day of menstrual cycle Hours after injection Endometrium Myometrium Endometriosis Adenomyosis Endometrioma Skeletal muscle Subcutaneous fat Plasma
estradiol
Monkey
Monkey 3, normal
2,
normal 1
1,103
3 1 1,014 603
Monkey 4, normal
14 28:
73;
242
291
Monkey 6, irradiated
2
15 if
;:
58 30 52
27
2
Comment
The present data indicate the high accumulation of radioactive estradiol in endometrial tissue in aberrant sites. This may be due to the presence of binding macromole-
Ave. at 2 hr.
2
394 394 417 20 51 36
Monkeys were injected with 1 fig of *H-estradiol per kilogram intravenously and radioactivity one or two hours later. Radioactivity is expressed as pFurie per milligram of wet tissue weight.
(Table II). The levels averaged as high as in the normal endometrium. All the radioactivity in endometriosis, adenomyosis, endometriomas, endometrium, and myometrium and 80 per cent from plasma co-chromatographed with authentic estradiol. The radioactivity in endometriosis, adenomyosis, endometrium, and myometrium is significantly higher (p < 0.05) than the radioactivity in skeletal muscle, subcutaneous fat, or plasma. The concentration gradients relative to plasma were endometriosis 11, adenomyosis 17, endometrium 16, and myometrium 8. The concentration of estradiol in endometriosis and adenomyosis was considerably higher (at least threefold) than the radioactive concentration in a wide variety of extrauterine tissues.15 In general, the radioactivity was higher in portions of endometriosis that were rich in glandular tissue and lower in portions containing substantial amounts of fibrous or fatty tissue.
Monkey 7, irradiated
9
652 639 77 33 120
129
SH-estradiol
Monkey 5, irradiated
543 908 64 57 61
in endometriosis
258 631 20 49 22 in
514 267 435 555 27 48 33
tissues was measured
cules with a high affinity for estradiol in endometriosis. In uteri of rats the accumulation in vivo is limited in capacity,16 and has a high specificity for drugs with estrogenic and antiestrogenic activity.l?> I8 This accumulation appears to be due to protein-containing macromolecules which tightly bind estradioLl@The first interaction of estradiol with the uterine protein appears to occur in the cytoplasm with the possiblesubsequenttransfer of the estrogen and cytoplasmic protein into the nucleus20 where it attaches to chromatin.21 The estrogen-protein macromolecule complex is thought to regulate gene activity promoting RNA synthesiswith subsequentincreasesin protein synthesis,tissuegrowth, and cellular division.22 The present finding of high accumulation of estradiol in endometriosis is compatible with the possibility that a similar sequence of biochemical events may be operative in endometriosis. We thank Mrs. R. S&none, Mr. Joseph Negri, and Miss Dorothy Butzko for their excellent
technical
assistance.
REFERENCES
1. Kliiver, Gynec. 2. Hartman, Gynec.
H., and Bartelmez, G. W.: Surg. Obstet. 92: 650, 1957. C. G.: Western J. Surg. Obstet. 52: 87, 1944.
3. 4. 5.
Pickering, McCann, OBSTET. Gardner,
D. E.: Folia primat. 8: 169, 1968. T. O., and Myers, R. E.: AMER. J. GYNEC. 106: 516, 1970. W. U., and van Wagenen, G.: IX
130
Eisenfeld, Gordner,
January 1, 1971 Amer. J. Obstet. Gynec.
and van Wogenen
International Cancer Congress, Tokyo, Abst. 0201, 1966. G., and Gardner, W. U.: In 6. van Wagenen, preparation. 7. Baird, Sir Dugald, Combined Textbook of Obstetrics and Gvnecoloav. Edinburgh. 1969, E. & S. Livingstone, Ltd., p. 821. 8. Kistner, R. W.: The Use of Progestins in Obstetrics and Gynecology, Chicago, 1969, Year Book Medical Publishers, Inc. 9. van Wagenen, G., and Morse, A. H.: Endocrinology 27: 268, 1940. 10. Te Linde, R. W., and Scott, R. B.: AMER. J. QBSTET. GYNEC. 60: 1147, 1950. 11. Scott, R. B., and Wharton, L. R.: AMER. J. OBSTET. GYNEC. 74: 852, 1957. 12. Jensen, E. V., and Jacobson, H. I.: Recent Progr. Hormone Res. 18: 387, 1962. 13. Eisenfeld, A. J., and Axelrod, J.: J. Pharmacol. Exp. Ther. 150: 469, 1965. -r,
-,
_
14. 15. 16. 17. 18. 19. 20.
21. 22.
Axelrod, L. R.: J. Biol. Chem. 201: 59, 1953. Eisenfeld, A. J., Gardner, W. U., and van Wagenen, G.: In preparation. Eisenfeld, A. J.: Biochim. Biophys. Acta 136: 498, 1967. Eisenfeld, A. J., and Axelrod, J.: Endocrinology 79: 38, 1966. Eisenfeld, A. J., and Axelrod, J.: Biochem. Pharmacol. 16: 1781, 1967, Toft, D., Shyamala, -G., and Gorski, J.: Proc. Nat. Acad. Sci., U.S.A. 57: 1740. 1967. Jensen, E. V.,. Suzuki, T., Kawashima, T., Stumpf, W., Jungblut, P., and DeSombre, E.: Proc. Nat. Acad- Sci., U.S.A. 50: 632, 1968. Maurer, H. R., and Chalklev. G. R.: YT. Molec. Biol. 27: 431, 1967. ” Hamilton, T. H.: Science 161: 649, 1968.