Testosterone

Carcinogenicity

Description of key information

All data presented are based in the result of a literature search (references are stated in the tables). For testosterone, only limited data on toxicity was available. Further data was found for testosterone propionate and other esters and used for classification.

Testosterone and its esters were tested in mice, rat and hamsters by subcutaneous injection or implantation, and in rabbits and baboons by intramuscular injection. It induced cervical-uterine tumors in mice and prostatic adenocarcinoma was induced in male rats.

The incidences of leukemia and of liver-cell and breast tumors in untreated mice of some strains were decreased by testosterone treatment, but the incidence of mammary tumors was increased by neonatal treatment of females of a mammary-tumor-virus-bearing strain. Neoplasm was induced in hamsters by a combination of estrogen and testosterone. Hyperplasia in the bladder, uteri and vaginal epithelium of rats were detected.

Justification for classification or non-classification

Due to the study results a carcinogenic effect of testosterone cannot be excluded. Additionally, it has to be taken into account that sex steroids in general can promote the growth of specific hormone dependent tissues and tumors.

Testosterone is classified as :

Category 3 (Xn, R40) according to Directive 67/548/EEC.

Category 2 according to Regulation (EC) 1272/2008 (CLP).

Additional information

All data presented are based in the result of a literature search (references are stated in the tables). For testosterone, only limited data on toxicity was available. Further data was found for testosterone propionate and other esters and used for classification. Although cancer test data are not required to fulfil the Annex VII obligations for a 1 -10 tpa registration dossier, summary data from the open literature are included here to provide support for classification conclusion. Read-across justification summary is attached to this endpoint summary.

Carcinogenicity-Animal Data

Test system	Test Substance	Application	Dose/Duration	Effect	Literature
C3H Mouse, male	Testosterone propionate (in sesame oil)	Subcutaneous	Once weekly for 25 (or more) weeks  0.25 – 1.25 mg/week	No hepatoma occurrence	Schenken & Burns, 1943 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
C3H Mouse, female	Testosterone propionate (in olive oil)	Subcutaneous	Once weekly for 12 months  1.00 - 1.25 mg/week	Reduced incidence of mammary tumors (3/12) compared to controls (18/38).	Jones, 1941 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Rabbit, female	Testosterone oenanthate (in sesam oil)	Intramuscular	15 mg on alternate weeks	2 Adenomatous polyps in 1/21 animals; no other findings	sark & Sommers, 1966 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Nb Rat, male	Testosterone propionate,	Subcutaneous implant	91 weeks 10, 20, 30 mg	Increased incidence for prostatic carcinoma at middle and high dose after about 40 weeks	Noble, 1977 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Wistar Rat, female	Testosterone together with nitrosamine in drinking water	Subcutaneous implant	50 mg (1:1 with cholesterol)	Enhanced incidence of bladder tumors	Okajima et al., 1975 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
A x C Rat, male, castrated	Testosterone propionate/cholesterol (1:3) together with N-2-fluoreenyldacetamide, 4 weeks,	Subcutaneous implant	48 weeks   200 mg/kg bw	Increased incidence of liver carcinomas	Reuber, 1976 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Hamster	Testosterone propionate and diethylstilboestrol	Subcutaneous implant	5 month interval testosterone propionate 100 diethylstilboestrol  25 mg	Induction of leiomyomas and leiomysarcomas	Rivière et al., 1961, Kirkman&Algard, 1965 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
BALB/cCrgl Mouse	Testosterone in water	Subcutaneous injection	First 5 days after birth  25 µg/d	Hyperplasic epithelial lesions, resembling epidermoid carcinomas (vaginal squamous-cell tumors)	Kimura&Nandi, 1967 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
BALB/cCrgl Mouse (MTV^+), female	Testosterone	Subcutaneous injection	First 5 days after birth   5 - 20 µg/d	Increased mammary tumor incidence after 8.5 months (mean)	Mori et al., 1976 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
SHN Mouse (MTV^+), female	5β-dihydrotestosterone in olive oil for the first 5 days after birth	Subcutaneous injection	200 µg/d	Increased mammary tumor incidence after 6.2 months (mean)	Yanai et al., 1977 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Albino Rat, female	Testosterone propionate in arachis oil	Subcutaneous injection	Weekly  0.5 mg , increased to 1 mg at the age of 21 days and to 2.5 mg at the age of 6 months	Theca-cell ovarian tumors, epithelial hyperplasia in the uteri	Horning 1958 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Sprague-Dawley Rat, female	Testosterone propionate in olive oil	Subcutaneous injection	Single dose on the day of birth 1.25 mg	Mammary tumors (fibroadenomas and adenocarcinomas)	Christacos et al., 1976 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Sprague-Dawley Rat, female	Testosterone propionate and DMBA	Subcutaneous injection	At day 2 after birth 1.25 mg   DMBA (20 mg, gavage) at day 50	Mammary dysplasia, no mammary tumors at day 300	Yoshida & Fukunishi, 1978 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Sprague-Dawley Rat, female	Testosterone propionate in sesame oil and DMBA	Subcutaneous injection	At day 2: 1.0  mg  DMBA (20 mg, gavage) at day 50	Increased auditory sebaceous gland tumors	Yoshida & Fukunishi, 1977 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Sprague-Dawley Rat, female	Testosterone and testosterone propionate in sesame oil and DMBA	Subcutaneous injection	Single dose at day 1 and 5 1.25 mg  DMBA (20 mg, gavage) at day 50	Little effect of 5 d treatment, reduces mammary adenocarcinoma but increased fibroadenoma at 1 d treatment. No tumors with testosterone alone.	Shellabarger & Soo, 1973 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
C57BL/Tw Mouse	1)Testosterone  2)Testosterone propionate  3) 5α-dihydrotestosterone  4) 5α-dihydrotestosterone propionate	Subcutaneous injection	Daily for the first 10 days after birth  1: 100 µg  2: 50 µg  3: 100 µg  4: 50 µg	Ovariectomized after 60 d. Vagina with stratified epithelium partly with superficial squamous metaplasia or cornification	Iguchi & Takasugi, 1976 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
C57BL/Tw Mouse	1) Testosterone  2) 5α-dihydrotestosterone	Subcutaneous injection	5 times (daily) from the day of birth  100 µg of either compound	Ovariectomized after 10 d.  Proliferation of the vaginal epithelium	Otha & Inoguchi, 1976 as cited by IARC. Monographs on the Evaluation of the Carcinogenic Risk of Chemicals to: World Health Organization, International Agency for Research on Cancer, 1972-PRESENT. (Multivolume work), V21 (1979)
Baboon Male	Testosterone Enenthate	Intramuscular injection,	200mg, once per week for 6 months.	Significant gravimetric and volumetric increases in the prostate; a definite glandular and marked stromal hyperplasia with fibrosis developed in the caudal lobe	Karr, JP et. al. Induction of benign prostatic hypertrophy in baboons., 1984. Urology: 23(3).
Rat	Estradiol plus testosterone	Implants killed at 8, 16 and 24 weeks after initiation.	Sampling of accessory sex glands for DNA adducts. Selective adduct formation in the dorsolateral prostate of rats treated with estradiol plus testosterone for 16 or 24 weeks.	Appearance coinciding with putative preneoplastic lesions and preceding carcinoma development	Han et al. 1995. Carcinogenesis 16(4), 951-954.
Mice; rabbits	Testosterone and its esters	Subcutaneous injection, or implantation or intramuscular injection	no data	Testosterone propionate implanted subcutaneously in mice induced cervical-uterine tumours, which metastasized in some cases; in rats, metastasizing prostatic adenocarcinomas were induced in males.   Neonatal treatment of female mice by subcutaneous injection of testosterone induced lesions of the genital tract and increased the mammary tumour incidence when the animals were adult. 5b-Dihydrotestosterone, which is considered to be hormonally inactive in adults, also increased the incidence of mammary tumours in mice when given neonatally by subcutaneous injections.	WHO. 1998. IARC Monographs on the Evaluation of Carcinogenic Risks to Humans, v21 Sex Hormones (II)
In vitro	testosterone and other steroids	2 mmol/L to DNA obtained from human surgical resections, rat liver, HepG2 cells, and calf thymus		no adducts were observed in DNA isolated from HepG2 cells incubated with 10-100 μmol/L testosterone for 24 h. The presence of a carbonyl group at C17 (which testosterone lacks) was predictive of DNA reactivity	Seraj, M.J., Umemoto, A., Tanaka, M., Kajikawa, A., Hamada, K. & Monden, Y. (1996) DNA adduct formation by hormonal steroids in vitro. Mutat. Res., 370, 49-59.In Food Who Food Additives Series: 43 Prepared by the Fifty-second meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA) World Health Organization, Geneva, 2000 IPCS - International Programme on Chemical Safety
Nobel Rats	Testosterone. Testosterone and estradiol	no data	no data	DNA strand breaks were observed in the dorsolateral prostate of Nobel rats treated with a combination of testosterone and estradiol but not in those treated with testosterone alone	Ho, S.M. & Roy, D. (1994) Sex hormone-induced nuclear DNA damage and lipid peroxidation in the dorsolateral prostates of Noble rats. Cancer Lett., 84, 155-162.. In Food Who Food Additives Series: 43 Prepared by the Fifty-second meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA) World Health Organization, Geneva, 2000 IPCS - International Programme on Chemical Safety

Carcinogencicity-Human

Description of Data	Literature
Increased testosterone levels may increase the risk for prostatic cancer. Limited evidence that anabolic steroids can cause both benign and malignant liver tumours	IARC Monographs, Suppl 7, 1987.
Epidemiological studies of the relationship between serum concentrations of androgens and estrogens and prostate cancer have yielded inconsistent but largely negative results (for references, see Dorgan et al., 1998; Ross et al., 1998). Since a possible association with markers of androgen metabolism (androstanediol glucuronide and DHT) has been observed in some studies, a complex, androgen-dependent, multi-gene model involving both androgen biosynthesis and metabolic pathways has been suggested to be critical for susceptibility to prostatic cancer	Ross, R.K., Pike, M.C., Coetzee, G.A., Reichardt, J.K.V., Yu, M.C., Feigelson, H., Stanczyk, F.Z., Kolonel, L.N. & Henderson, B.E. (1998) Androgen metabolism and prostate cancer: Establishing a model of genetic susceptibility. Cancer Res., 58, 4497-4505. And Dorgan, J.F., Albanes, D., Virtamo, J., Heinonen, O.P., Chandler, D.W., Galmarini, M., McShane, L.M., Barrett, M.J., Tangrea, J. & Taylor, P.R. (1998) Relationship of serum androgens and estrogens to prostate cancer risk: Results from a prospective study in Finland. Cancer Epidemiol. Biomarkers Prev., 7, 1069-1074. IN Food Who Food Additives Series: 43 Prepared by the Fifty-second meeting of the Joint FAO/WHO Expert Committee on Food Additives (JECFA) World Health Organization, Geneva, 2000 IPCS - International Programme on Chemical Safety