Structure and Origin of Uterine and Extragenital L=Ibroids Induced

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Structure and Origin of Uterine and Extragenital l=ibroids Induced Experimentally in the Guinea Pig by Prolonged Administration of Estrogens* Alexander Lipschotz, M.D., and Louis Vargas, Jr., M.D.

(From Department o/ Experimental Medicine, National Health Service o/the Republic o/Chile, Santiago, Chile) (Received for publication December 13, x94o)

The purpose of this communication is to present the These experimentally induced abdominal tumors findings of a detailed microscopical study of the sites present a smooth surface formed of a capsule com- of origin and stages of development of the subserous posed of flattened superficial cells (Plate 2, Figs. 2-A fibroid tumors induced in guinea pigs by prolonged and 2-B). The cells beneath the capsule resemble administration of estrogens. Details of treatment of fibroblasts. These cells have definite boundaries or the animals are given in the explanations of Plates I- 5. they are separated from each other by collagenous Subserous uterine tumors which can be induced in fibers (Plate 4, Fig. ix-C). guinea pigs by prolonged administration of estrogens, The masses of fibroid tumors arising from the apex as described by Nelson (26, 27), were found to be of the uterine horn may enclose the tubes or large fibroids. Lipschiitz, Iglesias, and Vargas (i3, 18, 22) tubal cysts. The demarcation between the muscular have shown that extragenital tumors in the abdominal coat of the tube and the tumor is not always sharp. cavity, induced by estrogens, also were fibroids. The In some instances, especially when the apical fibroid localization of these tumo~:s at various sites on the is small, the tumor is in close contact with an abun- uterus, pancreas, kidney, spleen, etc., have been de- dance of smooth muscle and adipose tissue (Plate 2, scribed by Iglesias (5), Vargas and Lipschiitz (32), Fig. 2-B), whereas the fibroid itself, to whose area the Bellolio (1), Murillo (25), Rodrlguez (3 o), and Lip- muscle tissue belongs, is covered with a capsule (Plate schlitz, Iglesias, and Vargas (15). Several examples of 2, Fig. 2-A). In some places fibrous tissue may also be uterine and extragenital fibroid tumors, induced in in direct contact with adipose tissue (Plate 4, Fig. guinea pigs treated with estradiol monobenzoate, are I I-A). Small epithelial tubes, probably belonging to shown in Plate i. We have demonstrated that similar the Wolffian body (Plate 2, Fig. 2-B) may be found fibroids at these sites can be induced by administra- surrounded by muscle tissue or connective tissue, as tion of all natural estrogens (estradiol, estrone, estriol) previously described by Lipschiitz (io). These fea- free or esterified (dipropionate, caprylate, benzoate- tures indicate that the apical fibroid is a tumor of the butyrate). Similar results were obtained by the ad- mesosalpinx. In the untreated normal animal patches ministration of such artificial estrogens as stilbestrol, in of compact smooth muscle may also be found near the the free form, by Lipschiitz and Vargas (I9), or esteri- tube. These masses of muscle probably hypertrophy fled, by Lipschiitz, Vargas, and Bruzzone (2i) or by under the influence of the estrogen, and are found hexestrol, by Lipschiitz and Egana (I2). subsequently in contact with, or incorporated in, the growing fibroid. GENERAL CHARACTERISTICS OF FIBROIDS Apical uterine fibroids occurred only in castrated The central portion of a typical large apical uterine females treated with estrogens. On the other hand fibroid is composed predominantly of fibrous tissue fibroids occurred at other sites in the abdominal cavity with few cells scattered between the bundles of fibers. in both normal and castrated guinea pigs. In some areas bundles of smooth muscles may be Abdominal fibroids can be induced experimentally found; in others the loose fibrous tissue may be edema- by estrogens both in noncastrated and castrated males. tous. These characteristics are shown in Plate 2, Figs. But larger quantities of estrogens administered over I-A, I-B, and I-C. longer periods of time are required to bring about * This investigation was aided by grants from The Ella Sachs these tumors in males than in females. The histologi- Plotz Foundation for the Advancement of Scientific Investigation, cal structure of the fibroids is the same in the males from The Jane Coffin Childs Memorial Fund for Medical Re- as in the females. Nevertheless, there is a pronounced search, from Mr. Adolfo Eastman (Limache, Chile), and by sex specificity in the collagenous tumoral reaction, as special Government funds for research work, by courtesy of the Minister of Health of the Republic of Chile. described by Koref, Lipschiitz, and Vargas (7), Jed- 236

Downloaded from cancerres.aacrjournals.org on October 2, 2021. © 1941 American Association for Cancer Research. Fibroid Tumors Induced in Guinea Pigs 237 licky, Lipschiitz, and Vargas (6), Chaume (i7), tumoral proliferation of the mesothelium of the serosa Szab6 (3i), and Palma (28). and of the endothelium of vascular and lymphatic In the castrated male a fibroid tumor is formed also spaces, or of the cells of the mesenchyme which are at the site of ligature of the spermatic cord. These in immediate contact with the endothelium. inguinal tumors, like the apical uterine tumors, may be In i938 LipscMitz, Vargas, and Iglesias (22) de- rich in smooth muscle. scribed an example of apparent proliferation of the As Lipschiitz (ii) has pointed out the structure of endothelium of vascular or lymphatic spaces in a fibroid the experimentally induced fibroid tumor of the male of the abdominal wall. In the past i8 months many or female guinea pig differs considerably from the additional observations have been made. Typical ex- spontaneous fibromyoma in women. In the experi- amples are shown in a series of sections of small mental fibromyoma there is much less order, particu- intramural fibroids of the uterus of a guinea pig larly in the arrangement of the fibrous tissue. The treated for 3 months with estrone (Plate 3, Figs. io-A compact myomatous tissue shows an orderly arrange- to Io-F). The cleft in the muscular septum between ment in both types of tumor. Another important dif- the uterine horns (Fig. Io-A) is lined with endo- ference is the abundance of fibroblasts near the capsule thelium and may be a lymphatic or vascular space. in the experimentally induced tumor (Plate 2, Fig. 2-A; A fibroid is in process of formation in the myometrium Plate 4, Figs. I I-B and r I-C). In addition numerous (Fig. Io-B). Neoformation of tumoral tissue here ap- cells of other types, some with pyknotic nuclei, may pears to be intimately related to the underlying cells be found in these fibroids (Plate 2, Fig. 3). of the mesenchyme and not with the endothelium. Small nodules or excrescences, called "tumoral seed" The club of tumoral tissue consists at its base of dense (Plate 2, Figs. 4-A and 4-B) deserve special attention. fibrous tissue and at its periphery of spindle cells, These have been found by Vargas and Lipschiitz (32) easily distinguishable from the muscle fibers of the on the inner abdominal wall, the surface of the stom- myometrium in which the fibroid is growing (Fig. ach, the spleen, and elsewhere. Their structure is io-C). The cells of the small nodule shown in Figs. similar to, though not always identical with, the typi- i o-D and i o-E are different from the spindle cells of cal experimental fibroid. Cells rich in cytoplasm may Fig. i o-C. These cells have the appearance of under- predominate in these small nodules (Plate 2, Fig. 5) going necrosis. In Fig. io-F definite evidence of necro- and an intimate relation with the mesothelial cells of sis is shown. the serosa is often evident. The structure of some of Another aspect of cellular proliferation is shown in these small nodules is the same as that of the large Plate 4, Fig. x I-A, from a splenic tumor. At the top fibroids. The cells at the periphery have the shape of the tumor is infiltrating the pancreas. But the contact fibroblasts and there is an abundance of fibrous tissue of the tumor with the pancreas is incomplete, so that in the center. In general these small excrescences seem a cleft remains between the two. There is abundant to have a strong tendency to undergo sclerotization, cellular proliferation in this cleft. These cells may be as shown in Plate 3, Figs. 6-A and 6-B. Although it is mistaken for proliferating endothelium; more prob- probable that the small excrescences may develop into ably they are the cells of the peripheral layer of the large fibroids it is not likely that every small nodule tumor, as shown also in Plate 4, Figs. I2-A and 14. or tumor seed of this type is potentially a large fibroid. One may find also an accumulation of cells sur- Apparently their development into large tumors is rounded by adipose tissue, as in Plate 4, Fig. i3, from often interrupted by sclerotization. a tumor of the hilum of the spleen. These accumula- tions are rich in blood vessels, as shown in Plate 4, ABUNDANCE OF CELLS Fig. I2-B, from an apical tumor. The processes re- sponsible for this accumulation of cells appear to be In our experimental animals there was often a re- intimately related to the blood vessels. One may as- markable augmentation of the mesothelium of the sume that the accumulation of cells around the blood serosa, as shown in Hate 3, Figs 7-A and 7-B. Al- vessels is due to proliferation of mesenchyme cells though the dii~erence between the serosal cells of underlying the endothelium, but the question is far normal guinea pigs and guinea pigs under treatment from being settled. with estrogens is not always conspicuous, nevertheless in some instances there is a striking accumulation of INFILTRATION cells rich in cytoplasm on the surface of the tumor. The importance of these findings, however, should not The extra-uterine fibroids invade certain tissues. The be overestimated. A similar transformation of the behavior is different according to the organ. A fibroid mesothelium of the serosa may occur as a result of in close contact with the kidney or the spleen is always inflammation of the peritoneum. The findings raise separated from these structures by a sharp line; i.e., the relevant question whether there is in these cases a by the fibrous capsule of the organ which may be very

much thickened at the line of contact. On the other estrogens, present all transitions from fibromyoma to hand, other organs, such as the pancreas, liver, smooth fbroid, sometimes with an abundance of cells. As to muscle, and striated muscle are infiltrated by the tu- their origin it is very likely that proliferation starts moral tissue. There is a disintegration and destruction at different points of the serosa and in the proximity of lobules of the pancreas which are in contact with of lymphatic or vascular spaces. It is as yet not possible the tumor or are surrounded by it (Plate 5, Fig. I5). to say whether the celomic, lymphatic, or vascular In the same way, though in a lesser degree, liver tissue endothelium becomes active or whether only cells un- suffers disintegration when in contact with the tumor derlying the endothelium enter into proliferation. The (Plate 5, Fig. I6). latter is more probable than the former. There can be Infiltration of smooth muscle tissue has been studied no doubt that cells of the mesenchyme become acti- in sections of the stomach and various parts of the vated so as to proliferate abundantly, under the in- intestine (Plate 5, Fig. 18). Intermingling of smooth fluence of a prolonged treatment with estrogens. There muscle tissue of tumor and myometrium may occur is an accumulation of cells around the blood vessels also (Plate 5, Fig. I7). In general one may be in even outside the fibroid, as in adipose tissue. The doubt as to how to interpret the condition; i.e., eventual results of these proliferative processes are, whether there is only infiltration of the muscle tissue though probably not always, spindle-shaped cells, by the tumor or whether the muscle tissue participates smooth muscle fibers, and collagenous fibers. itself in the proliferation. It is the same in the male It is natural to question whether these experimental with the vas deferens when surrounded by a tumor. abdominal fibroids can be compared with the uterine The invasion and disintegration of striated muscle by fibroids of the woman. There are certain structural tumors of the abdominal wall (Plate 5, Fig. I9) or of differences which we have already insisted upon. the diaphragm is also very remarkable. These differences are not coincident with species dif- The resistance of certain organs as kidney and spleen ferences as shown by the fact that the only spontane- against invasion is all the more remarkable as their ous uterine fibromyoma in the guinea pig which Lip- surface and hilum are so often the site of fibroids of schiitz (I1) had the opportunity to observe was considerable size. structurally very similar to the spontaneous fibroids of the woman and very different from the experimental (}ENERAL FIBROSIS fibroids of the guinea pig. The great abundance of cells in some of the experimental tumors and the great Fibrous strands on the inner abdominal wall, in the tendency to invade certain organs has also to be men- mesentery, and other places often accompany fibroids tioned here. or they are the only manifestation of the collagenous But one should not attribute too much importance to proliferative reaction as reported by Vargas and Lip- these structural and biological differences between ex- schlitz (32). These strands are very poor in cells. perimental and spontaneous fibroids. One must not Lipschfitz and Vargas (17) found accidentally also a forget that in these experiments the tumor is induced fibrous reaction around the glands of Brunner in the under quantitative and time conditions which are duodenum. The thickening of the capsule of the probably very different from those in the body when kidney (Plate 3, Fig. 8-B) or of the uterus (Plate 3, a fibroid originates spontaneously. Quantities of estra- Fig. 9) may be extensive. In the latter case there is in diol no less than ioo to I5o times greater than the some places an abundance of smooth muscle fibers in hysterotrophic or physiological dose are necessary to the capsule itself. The thickening of the capsule of the induce experimental fibroids. With esterified estradiol kidney is sometimes noticeable even to the naked the difference between the hysterotrophic and the eye because of the whitish color of the surface. There tumorigenic dose is smaller than with the free hormone is also a rich development of fibrous tissue in the mam- (16, 3 o, 2o), but here again we must insist that es- mary gland, sometimes, though very rarely, with for- terification overthrows completely the chronological mation of adenofibroma. conditions of the action of ovarian estrogens. No It is well known that the stroma of the prostate in- knowledge exists as yet on these fundamental quanti- creases greatly under the influence of a prolonged tative and chronological questions in spontaneous tu- treatment with estrogens, as reported by Parkes and morigenesis and one cannot expect for the moment to Zuckerman (2 9), and De Jongh and associates (3). induce experimental fibroids structurally identical with Loeb, Suntzeff, and Burns (:24) have described the the spontaneous ones. influence of estrogens on the stroma of other organs. The question has been raised whether the fibromyoma, experimental or spontaneous, is a neoplasm at DISCUSSION all (8, 9)- If autonomous growth; i.e., ability to grow From the observations reported in this paper, it is even when the stimulus that has led to its appearance evident abdominal tumors, experimentally induced by is withdrawn (9), is an essential of true neoplasm,

benign or malignant, then spontaneous fibromyoma tO infltrate and destroy certain differentiated tissues is not a tumor; fibromyoma in the woman ceases grow- such as pancreas, liver, striated muscle, and smooth ing and begins to regress when the ovary ceases its muscle. endocrine function. There is no tumoral cell, or tumoral autonomy of spontaneous fibromyoma (29). REFERENCES The experimental fibromyoma also begins to regress i. BEELOmO, P. Estudio comparativo sobre la acci6n histero- when treatment is suspended and important structural tr6fica y tumorlgena del benzoato y dipropionato de changes take place (14). These changes, hyalinization estradiol. Public. Med. Exper. (Chile), 2:1939. and ossification, are similar to those in the fibromyoma 2. CHAVME, J. Nuevos estudios experinaentales sobre la espe- of the woman in the menopause. cificidad sexual en la reaccidn tumoral provocada por substancias estrdgenas. Rev. Chile de hig. y reed. prevent., We were also unable to obtain permanent grafts of in press. our experimental fibroids though they may persist for 3. DE JONGH, S. E., D. J. KoK, and L. A. VAN DER WOERD. some time in the host into which they have been trans- Paradoxe Wirkungen des Follikelhormons bei m~innlichen planted. Tieren II; die Beeinflussbarkeit durch gonadotropes Hor- mon; die Beziehungen zur Prostatahypertrophie. Arch. The question whether the fibromyoma, spontaneous Internat. de pharmacodyn, et de th6rap., 58:31o-331. or experimental, is a true neoplasm can be settled 1938. only on the broader basis of experimental tumorigene- 4. GARDNER,W. U., E. ALLEN, G. M. SMITH, and L. C. STRONG. sis. If the fibromyoma is not a neoplasm, then neither Carcinoma of the Cervix of Mice Receiving Estrogens. J. A. M. A., IIO:II82-II83. I938. is the uterine adenoma, since the latter also can be 5. IGLESIAS, R. Tumores experimentales uterinos y extrageni- produced by injection of follicular hormones even with tales provocados por el benzoato de estradiol. Public. Med. relatively small quantities (23). These probably will Exper. (Chile), I:I938. regress partially when treatment is suspended. But 6. JEDLICKY, A., A. LIPSCH/SWZ, and L. VARGAS vmS. La on the other hand, atypical epithelial growth induced sp&ificit~ sexuelle dans la rdaction tumorale conjonctive vis-a-vis de l'hormone folliculaire (Caprylate et I7-ben- in the uterus by prolonged administration of estrogens zoate-3-n-butyrate d'oestradiol). Compt. rend. Soc. tie can eventually persist when administration is sus- biol., 1,:10:1466-1469. 1939. pended (14) and can even become transplantable as 7. KOREV, O., A. LWSCHOTZ, and L. VARGAS rms. Sp&ificit6 shown by Gardner, Allen, Smith, and Strong (4)" sexuelle et tumorigen~se. Compt. rend. Soc. de biol., There is evidently a transition from proliferating epi- 1`3o:3o3-3o6. 1939. 8. LANCET (Leading Article). The Origin of Fibroids. 9:1329- thelial cells without autonomous growth, to true tu- 133 ~ 9I939. moral epithelial cells. This transition is the funda- 9. LANCET (Annotation). The Prevention of Fibroids. 2:438. mental problem of malignant neoplasms and of ex- 1939. perimental tumorigenesis in general. lO. LxPscn/Sxz, A. Fibromioma uterino como slntoma de endo- crinopatia, desde el punto de vista experimental. Primer One may also question whether the great capacity Congreso Chileno y American() de Cirugia, Santiago of the experimental fibroid to invade certain neighbor- (Chile), 252-282. 1939. ing tissues, which we have described, indicates transi- I I. LIPSCH/JTZ, A. Arch. Path, in press. tion from benign growth to malignancy. One should l Z. LiPscH/SWz,A., and E. EGA~qa. Unpublished data. indeed keep in mind that a similar behavior of con- 13. LmSCH/STZ, A., and R. IGEESIAS. Multiples tumeurs ut6rines et extrag~nitales provoqu6es par le benzoate d'oestradiol. nective tissue can be seen also in other conditions of CDmpt. rend. Soc. de biol., I29:519-524. 1938. fibroblastic proliferation which has nothing to do with 14. LIPSCHi)TZ, A., R. IGLESIAS, and L. VARGASVIES. R~gression tumorigenesis, as in hepatic disease. But have we not du fibromyome exp6rimental, et r6sistance des formations overestimated the gap between localized tumorigenesis tumorales 6pith61iales, dans la carence hormonale follicu- and general tissue reactions? Our own results with laire. Compt. rend. Soc. de biol., 1,3o:1536-154o. 1939. I 5. LIPSCH/]ITZ,A., R. IGLESIAS, and L. VARGAS, JUN. Proc. Soc. experimental extra-uterine fibroids may serve as a first Exper. Biol. & Med., in press. step in dealing with this question. 16. LIPSCH/STZ, A., F. RODRIGUEZ, and L. VARGAS FINS. Dose hyst6rotrophe "utile" et dose "tumorig~ne" de t'oestradiot SUMMARY et de l'oestrone libres. Compt. rend. Soc. de biol., I,3O: 939-942 9I939. The microscopic structure of experimental fibroids, 17. LIVSCH/STZ,A., and L. VARGASVIES. Etude comparative sur uterine and extra-uterine, induced by a prolonged l'action tumorig~ne de diffdrentes substances oestrog~nes. Compt. rend. Soc. de biol., I,3o:9-11. I939. administration of estrogens, is described. I8. LIPSCHUTZ,A., and L. VARGAS, JUN. Experimental Tumori- The participation of the cellular elements of the ab- genesis with Subcutaneous Tablets of Oestradiol. Lancet, dominal wall and of those of lymphatic and vascular I:x313-1318. 1939. spaces in the proliferative process is discussed. 19. L1PSCH/JTZ, A., and L. VARGAS, ]UN. Tumorigenic Powers The differences between experimental and spontane- of Stilbcestrol and Follicular Hormones. Lancet, 1:541- 543. 194o- ous fibroids are emphasized, especially the abundance 20. LlPSCHi)TZ, A., L. VARGAS, JUN., H. BAEZA-ROsALES, and H. of cells in the experimental tumor and their capacity BAEZA-HERRERA. In press.

21. LIPSCHi3TZ, A., L. VARGAS, ]UN., and S. BRUZZONE. Not yet of the Guinea Pig by the Prolonged Administration of published. Oestrogenic Hormone. Anat. Rec., 68:99-1o2. 1937. 22. Ln~SCHOTZ, A., L. VARGAS FILS, and R. IGLESlAS. Sur la 27. NELSON, W. O. Atypical Uterine Growths Produced by Pro- structure microscopique des tumeurs ut6rines et abdomi- longed Administration of Estrogenic Hormones. Endo- nales dues a l'action du benzoate d'oestradiol. Compt. crinology, ~4:5o-54. 1939. rend. Soc. de biol., I29:524-528. 1938. 28. PALMA,J. Especificidad sexual en la reacci6n tumoral provo- 23. LIPSCHf.)TZ, A., L. VARGAS, JUN., A. ]EDLICKY, and P. BEL- cada por estr6genos artificiales. Public. Med. Exper. LOLIO. The Minimum Quantity of Estrogen Required to (Chile), 6:1-27. I94o. Induce Atypical Epithelial Growth of the Uterine Mucosa 29. PARKES, A. S., and S. ZUCKERMANN. Experimental Hyper- plasia of the Prostate. Lancet, I:925. 1935. in the Guinea-Pig. Am. I. Cancer,/19:185-198. 194o. 30. RODRiGUEZ, F. Estudio experimental comparativo sobre la 24. LOEB, L., V. SUNTZEFF, and E. L. BURNS. The Effects of dosis histerotr6fica fitil y la dosis tumorlgena de las Age and Estrogen on the Stroma of Vagina, Cervix and hormonas foliculares libres (Estradiol y estrona). Public. Uterus in the Mouse. Science, 88:432-433. i938. Med. Exper. (Chile), 5:194o. 25. MURILLO, R. La acci6n protectora del ovario en la tumori- 3 I. SZAB6, I. Tumorig6nesis conjuntiva provocada por sub- g6nesis conjuntiva experimental y el papel de la proges- stancias estr6genas y la vitamina. Rev. Chile de hig. y terona en esta acci6n. Public. Med. Exper. (Chile), 4: reed. prevent, in press. 194 ~ 9 3 2. VARGAS, L., FXLS, and A. LXPSCHi3TZ. Sur la gen&e des 26. NELSON, W. O. Endometrial and Myometrial Changes, In- tumeurs conjonctives provoqu6es par le benzoate d'oestra- cluding Fibromyomatous Nodules, Induced in the Uterus diol. Compt. rend. Soc. de biol., I29:8IO-813. 1938.

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Fro. XIII. 34- Large bilateral tumors of the mcsometrium: o17 mcsocolon and largc minor of thc duodenum near tile fibr~us masscs in the apical rcgion of thc right uterine horn: pylorus, l.owcr left, several largc tumors at thc hilum of the apical tumor of the left uterinc horn united with tumor of spleen. Lower right, right kidncy with subserous fibroid ncar the hilum of thc splecn, involving also thepancrcas. Thelcft the lower polc and subserous tumor of the abdominal wall. kidney is displaced to the right by pressure of masses of Mag. X 1.5. tumor. Castrated fcmalc guinea pig which receivcd 25 injec- Fie,. XIII. 41. Large fibroid tumor between spleen, pan- tions of 8o #gin. of estradiol monobenzoate in 6.3 days. creas, and stomach. The tumor has been cut across to show Mag. )< 1.5. the diameter in the mid-region. Pedunculated uterine Fro. XIII. 38. Fibroid tumors from castrated female guinea fbroid near the parametrium. Castrated female guinea pig pig treated in same manner as XIII. 34. Upper left, tumor treated as XIII. 34. Mag. X 1.5.

PLATE 2

Fins. I-A, I-B, and i-C. Sections from a large apical Fro. 3- Section from a fibroid between the spleen and tumor of the left uterine horn, extending to the kidney and stomach in female guinea pig, II. 38, injected during 8I days spleen and penetrating the pancreas. From castrated female with doses of 80 /,gm. estradiol benzoate (total amount 2.6 guinea pig, II. 14, injected thrice weekly for I22 days with mgm.), showing abundance of cells of various types sur- 8o /*gm. estradiol benzoate (total 3.9 mgm.). Van Gieson rounded by fibrous tissue. Van Gieson stain. Mag. X 560. stain. Mag. X 2oo. Fins. 4-A, 4-B, and 4-C. Sections from small nodules, Fro. I-A. Loose and dense fibrous tissue in great disorder; "tumoral seed," on the serosal surface of the stomach in cells scattered between the fibers. castrated female guinea pig, II. 17, injected with doses of FIG. I-B. Bundles of smooth muscle fibers and fibrous 80 /~gm. estradiol benzoate during 68 days (total amount tissue. 2.2 mgm.). Van Gieson stain. Fro. I-C. Edematous fibrous tissue. FIG. 4-A. Abundant fibrous tissue between the tumor and FIGS. 2-A and 2-B.--Sections from a tumor between the muscle layer; penetration of fibrous tissue between muscle spleen and pancreas in castrated female guinea pig, II. IS, fibers. Mag. X 23. injected with doses of 8o /~gm. estradiol benzoate (total FIG. 4-B. Two small nodules of "tumoral seed" on the amount 3.9 mgm.) during I22 days. Van Gieson stain. serosal surface of the abdominal wall. Mag. X io. FIG. 2-a. Portion of the tumor near the spleen, showing FIG. 4-C. Small cellular fibroid on the spleen, surrounded capsule consisting of flattened cells, fibroblasts (?) beneath by adipose tissue. Mag. X 45- the capsule and other cells separated by fibrous tissue. Mag. FIG. 5. Section of small tumors on the spleen in a cas- X 200. trated male guinea pig, VIII. I5, injected with doses of FIG. 2-B. Portion of the tumor near the apex of the uterine 8o/,gin. of estradiol benzoate-butyrate during i o4 days (total horn, showing smooth muscle tissue in contact with adipose amount 3-5 mgm.). Abundance of cells. Van Gieson stain. tissue and a Wolflian tubercle (?). Mag. X 45. Mag. X 2oo.

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PLATE 2

PLATE 3 FIGS. 6-A and 6-B. Section of nodule of tumoral seed on FIGS Io-A, I0-B, Io-C, Io-D, Io-E, and io-F. Sections of the tunica albuginea of the testis in male guinea pig, VIII. 9, small tumors in the uterine wall of castrated female guinea injected with doses of 8o /~gm. of estradiol benzoate-butyrate pig, V. 28, injected with doses of 4oo /zgm. of free estradiol during 99 days (total amount 3.3 mgm.). The nodule is poor during I23 days (total amount 2i mgm.); same animal as in in cells, does not contain muscle fibers and shows sclerotiza- Fig. 9- Van Gieson stain. tion. Van Gieson stain. Mag. X 2oo. FIG. I o-A. Muscular septum between uterine horns con- FiGs. 7-A and 7-B. Sections of spleen to show serosal sur- taining glandular cysts, showing blood-containing clefts in face. Van Gieson stain. the muscle tissue, with 2 fibroids. Mag. X Io. FIG. 7-A. Mesothelium on surface of the spleen in a FIG. Io-B. Small fibroid lying in a cleft in the muscle normal female guinea pig, C.u. I3, weight 555 gm. tissue. The cleft is lined with endothelium. Dense fibrous FIG. 7-B. Thickened mesothelial covering of spleen in male tissue at the base of the fibroid. Mag. X 45- guinea pig, VIII. 15, treated as described in legend for Fig. 5. FIG. Io-C. Small fibroid in the myometrium. The cleft is Mag. X 2o0. lined by endothelium. Spindle-shaped cells at the periphery FIGS. 8-A and 8-B. Sections through capsule of kidney. of the fibroid, showing a sharp contrast between the tumor Van Gieson stain. Mag. X 2oo. cells above the cleft and the muscle cells below the cleft. FIG. 8-A. Thin capsule in normal animal C.u. 13. Mag. X 200. FIG. 8-B. Thickened capsule of kidney in male guinea FIG. I o-D. Cleft in the myometrium with small tumors pig, VIII. 6, injected with doses of 80 #gm. estradiol benzo- separated from the muscle tissue by fibrous tissue. Mag. ate-butyrate during 87 days (total amount 2.8 mgm.). X 45. FIG. 9. Section through thickened serosal surface of the FIG. io-E. The same nodule showing difference between uterus in castrated female guinea pig, V. 28, injected with the apparently necrotic cells (above) and the cells of the doses of 4o0 /zgm. of free estradiol during 123 days (total myometrium (below). Mag. X 20o. amount 2i mgm.). Thick layer of smooth muscle fibers FIG. Io-F. Same cleft as in Figs. xo-D and io-E, showing beneath the mesothelium; dense fibrous tissue between this necrosis and accumulation of pigment in the tumor nodule. layer and the myometrium. Van Gieson stain. Mag. X 200. Mag. X 45.

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PLATE 4 Fins. II-A, I I-B, and II-C. Sections of fibroid between tissue and numerous blood vessels. Different types of cells the spleen and pancreas in castrated female guinea pig, II. may be due to proliferation of mesenchymal cells around the St. 2, injected with doses of Ioo /zgm. of stibestrol during capillaries. Mag. X 56o. Io5 days (total amount 4.3 gm.). Van Gieson stain. Fie. I3. Accumulation of cells rich in cytoplasm in FIG. I I-A. Cells proliferating in the cleft between the adipose tissue near the hilum of the spleen in same animal tumor and the pancreas. Mag. X Ioo. as in Fig. 5- Mag. X 2oo. Fro. I I-B. Cells rich in cytoplasm on the surface of the Fro. I4-A and I4-B. Sections through cleft between a tumor, with fibroblasts below the surface. Mag. X 200. tumor of the mesentery (right) and the intestinal wall Fro. x I-C. Fibroblasts. Mag. X 45 o. (left) in castrated female guinea pig, I. 3, injected with FIc. I2-A and I2-B. Section of apical tumor reaching the doses of 40 /zgm. estradiol benzoate during I2I clays spleen in castrated female guinea pig, VI. 38, injected with (total amount 2 mgm.). At other places the tumor adhered doses of 80 /zgm. dipropionate during 9o days (total amount to the intestine and infiltrated the muscular coat. Van 3.2 mgm.). Van Gieson stain. Gieson stain. Mag. X 240. Flc. I2-A. Cells rich in cytoplasm around a cleft (vascular FIc. I4-A. Tip of nodule composed of spindle-shaped cells. space ?) in the center of the fibroid. Mag. X 3 oo. F1c. I4-B. The nodule in contact with the intestinal wall. Fie. x2-B. Accumulation of cells in contact with adipose

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PLATE 4

PLATE 5

FXG. 15. Section of apical tumor infiltrating the pancreas, Fro. I8. Section showing infiltration of the muscular coat from same animal as in Fig. I-A, showing clusters of pan- of the rectum by fibrous tissue of a pelvic tumor in cas- creatic cells surrounded by fibrous strands of the tumor, trated female guinea pig, I. I9, injected with 4 ~ /~gm. of causing disintegration and destruction of glandular tissue. estradiol benzoate during I25 days (total amount 2.1 Van Gieson. Mag. )< I25. mgm.). Hematoxylin and eosin stain. Mag. )< 45. Fro. 16. Section of subserous fibroid of the liver in cas- Fins. 19-A and I9-B. Sections of striated muscle infiltrated female guinea pig, II. 21, injected with doses of trated by fibrous tissue of a large fibromyoma in contact 8o /*gm. estradiol benzoate during 8o days (total amount with the abdominal wall, in castrated female guinea pig, 26 mgm.). No sharp demarcation between the tumor and II. I4, same animal as in Fig. i. Mag. )< too. the liver. Van Gieson stain. Mag. X 56o. FIG. I9-A. Muscle fibers in transverse section surrounded FIG. 17. Section of subserous tumor of uterus in castrated by abundant fibrous tissue. Van Gieson stain. female guinea pig, V. 28, same animal as in Figs. 9 and Io, FIa. I9-B. Muscle fibers in longitudinal section separated showing intermingling of muscle fibers of tumor and hyper- by fibrous tissue. Hematoxylin and eosin stain. trophied external layer of the myometrium. Van Gieson stain. Mag. )< 45.

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Downloaded from cancerres.aacrjournals.org on October 2, 2021. © 1941 American Association for Cancer Research. Structure and Origin of Uterine and Extragenital Fibroids Induced Experimentally in the Guinea Pig by Prolonged Administration of Estrogens

Alexander Lipschütz and Louis Vargas, Jr.

Cancer Res 1941;1:236-249.

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