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The Fibromatogenic Action of Specific Urinary Estro- Gens (Metahormones) in the Guinea Pig*T

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The Fibromatogenic Action of Specific Urinary Estro- Gens (Metahormones) in the Guinea Pig*T The Fibromatogenic Action of Specific Urinary Estro- gens (Metahormones) in the Guinea Pig*t Alexander LipschCitz, M.D., 1Ren~ Thibaut, M.D., and Luis Vargas, Jr., M.D. (From the Department o/ Experin',er~tal Medicine, National Health Service o/ the Republic o~ Chile, Santiago, Chile) (Received for publication September i7, 194~) Conversion of estradiol and estrone, or ortho- and the human placenta, as shown by Doisy (2) and Mar- parahorlnones, ~ into estriol, a specific urinary meta- rian (i9). Since both are present in the ovary (2), hormone of women, acquires a new interest since, in they can be considered as primary ovarian estrogens. guinea pigs, toxic phenomena, including tumorigenesis, Estriol and equilenin were used as representatives of are elicited by prolonged action of follicular hormones. urinary estrogens. The first is present also in the Further interest is added by the findings of Pincus placenta, but both are absent from the ovary. They can and Graubard (2o) who have reported that cancerous be considered as specific urinary estrogens. women appear to be unable to convert estrone into Pellets of crystalline hormones were prepared by estriol to any appreciable extent. It is generally as- compression in a suitable hand press as described in sumed that this conversion signifies inactivation; i.e., detail in the thesis of Thibaut (23). The weighed that substances of high estrogenic activity (estradiol pellets were implanted subcutaneously in castrated and estrone) are transformed into substances of low guinea pigs. After the death of the animals the pellets estrogenic activity (estriol in the urine of the pregnant were recovered and weighed again. The animals were woman; different urinary estrogens of the mare). One killed and autopsied at different intervals after im- may also assume, tentatively, that by means of this plantation of the pellet. The fibrous reaction was conversion the body protects itself against the toxic or classified in accordance with criteria specified in pre- tumorigenic action of follicular hormones. This as- vious papers by Lipschiitz and Vargas (i4) , and Lip- sumption was tested by making a quantitative com- schiitz, Bellolio, Chaume, and Vargas (io). The re- parison of the faculty of different ovarian and urinary sults with respect to absorption and total tumoral effect estrogens to elicit abdominal serosal fibroids in the as obtained in lI 4 animals are summarized in Table I guinea pig. and Fig. i. MATERIALS AND METHODS COMPARATIVE FIBROI~'IATOGENIC ACTIVITY OF DIFFERENT ESTROGENS Alpha-estradiol and estrone were used as represen- tatives of ovarian estrogens. Both are found also in the All of the four estrogens were tumorigenic. The urine, especially estrone, which can be extracted from localization of tumors was the same as in animals receiving estradiol and its esters, or artificial estrogens, * This investigation was aided by grants from Thc Jane Coffin as reported previously by Lipschiitz and Iglesias (io), Childs Memorial Fund for Medical Research, from The Rocke- feller Foundation, and from Mr. Adolfo Eastman of Limache, Iglesias (6), Lipschiitz, Iglesias, and Vargas (ii), and Chile. These grants were administered by Professor A. Lipschfitz. Szabo (22). However, there were considerable differ- Crystalline estradiol was supplied by Dr. Carl Miescher, of the ences as to the degree of the reaction. firm of Ciba in Basel. Estrone, estriol, and equilenin were sup- plied by Dr. Oliver Kamm, of Parke, Davis & Co. The first manifestations were found as early as 2~ j" The senior author is responsible for the interpretation of to 3 ~ days after implantation of pellets. These were results and preparation of the manuscript. small, barely visible, nodules on the spleen, the stom- i In recent years Lipschfitz (8) has used a new Spanish ach, and sometimes at the junction between the uterus terminology to indicate the physiological relationships of the and the parametrium. These primary manifestations different sex hormones. According to this terminology estradiol, progesterone, and testosterone are orthohormones; estriol and the of a fibrous reaction graded as o. 5 were present with specific urinary estrogens of the mare, pregnandiol, androsterone, all four estrogens. and dehydroandrosterone are metahormones. The term para- The tumoral reaction increased with time. At 8o hormones may be appropriate for those hormones whose physi- and ~2o days marked differences became evident be- ()logical activity is doubtful; for example, estrone. The term prehormone also may be useful. It must be left to others to tween estradiol and estrone on one hand, and estriol judge whether this terminology is convenient in English.-- and equilenin on the other. As noted in our previous Authors' note. reports (6, I~, I5) , individual variations were con- 45 Downloaded from cancerres.aacrjournals.org on September 28, 2021. © 1942 American Association for Cancer Research. 46 Cancer Research siderable. Resistant animals were present in the groups it is known that estrone is absorbed more slowly than given each of the four estrogens. Nevertheless, it was estradiol. In our experiments, results of which are a striking finding that, in experiments continued for shown graphically in Figs. i and 2, we found that 5 ~ days or longer, 6 out of 33 animals treated with the absorption of estrone was considerably slower than estradiol and estrone developed high tumoral reactions, that of estriol or equilenin. Nevertheless, the tumori- "FABLE l: TUNIORALEFFECT IN 71 CASTRATED FFMALE GUINEA Pros (2IO TO 53 ~ GM. WIIEN RECEIVING THE IhiPLANT) WITH SUBCUTANEOUSLYIMPLANTED PELLETs OF FOUR DIFFERENT ESTROGENS Number of animals with Average total tumoral Duration of absorption Total tumoral effect effect not experiment Number of per (lay c ~" , less than Estrogen in days animals #gm. Average Range grade 6 Estradiol .................. 5 ~ I o 75 2.7 o-9 I ] Estradiol .................. 80 5 I8 4.6 oq2 i[ 6 Estrone ................... 50 5 I3 3.2 o-5-8 I out of Estrone ................... 80 5 I I 3.9 o-5-6.5 I / 33 Estrone ................... I2O 8 5.5 4.6 1.5-I2 2j Estriol .................... 50 4 44 ,.9 ,,-5 Estriol .................... 80 5 20 2. 4 0.5-4 Estriol ................... i2o 8 15 3.1 0-8 1 I out of Equilenin ................ 50 5 22 0.5 o-I.5 o| 38 Equilenin ................. 80 9 I9 0.7 0-4 o ] Equilenin .................. I 2o 7 15 o .... - I o ,17 0 Estradiol l&l,5 i0 20 Es ~rone /~ 30 ~ I 40 t J -~ 1 ~ i ~ Equilenin 50 %-- / Estrio~ i #/ 60 . I ,. ~. Estriol 70 1 5 i 80 I ~iuilenin ]!8 90 2 Days IDO [Days L I 21 30 50 80 120 21 30 50 80 12( Fro. I.--Graphs showing degree of fibromatogcnic action of estradio], estrone, estriol, and cquilenin in i I4 castrated guinea pigs. The estrogens in pellets were implanted subcutaneously. Each point is the average of results in 4 to Io animals. The num- bers near each point indicate the average absorption in mgm. The fibromatogenic action of estriol and equilenin is considerably less than that produced by equal or smaller quantities of estrone or estradiol. For details see Table I and Thibaut (23). FIG. 2.--Graphs showing percentage absorption from subcutaneously implanted pellets of estradiol, estrone, estriol, and equilenin in 114 female guinea pigs. All pellets were of the same diameter, 1.6 ram., but not of the same length. The weights of the pellets varied between z.~ and 6.2 mgm. Rate of absorption of estrone was less than that of other estrogens. For details see Thibaut (23). not lower than grade 6, while among 38 animals genic action was enormously superior with estrone. treated with estriol and equilenin only one developed Our results show, therefore, that specific urinary estro- such a reaction, the tumoral reaction generally failing gens are less fibromatogenic than ovarian estrogens. to appear. This confirms the statement of Lacassagne (7) who The differences in the degree of tumoral reaction found equilenin to be less active than estrone in were not due to the differences in the quantities of eliciting mammary adenocarcinoma in mice, although hormones absorbed. From the work of Deanesly (1) no exact quantitative data on this point are available. Downloaded from cancerres.aacrjournals.org on September 28, 2021. © 1942 American Association for Cancer Research. L ipschiitz et al.--Fibromatogenic Action of Urinary Estrogens 47 COMPARATIVE HYSTEROTROPIC ACTION the formation of polyps, may be obtained also with equilenin. Genital bleeding occurred with equilenin The question arises as to whether the lower fibro- as with other estrogens. matogenic action of certain estrogens is concomitant with a lesser estrogenic activity. According to general opinion this should be so because, in the Allen-Doisy COMPARATIVE FIBROMATOGENIC ACTION OF test, estriol and the urinary estrogens of the mare are FOLLICULAR HORMONES ABSORBED FROM less active than estradiol or estrone. The same results PELLETS AND INJECTIONS are obtained with the uterine tests. When rats are In previous experiments we (13 , 21) found that 38 injected once daily for 5 days with estrogens in oil, to 5 ~ injections of 75o to 2oo /,gm. of estradiol, or the highest uterine ratio (ratio of the weight of the ~5 o to 3o0 /,gin. of estrone, in oily solution; i.e., a uterus in mgm. to body weight in gm.) is obtained with estradiol. With estrone this ratio is somewhat less and with estriol and equilenin it is considerably below the value with estradiol, as shown by the work of Dorfman (3)- Similar statements concerning this ratio in the mouse were made by Evans, Varney, and Koch (5)- Emmens (4), however, discovered that the / Estrone differences in the degree of estrogenic effect exerted Estriol by the natural estrogens, as evidenced by cornification of the vaginal mucosa, depend in part upon the num- d ber of injections made into the same animal for pur- pose of assay.
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