Profile of Histopathological Type and Molecular Subtypes of Mammary Cancer of DMBA-induced Rat and its Relevancy to Human Breast Cancer


  • Ika Fidianingsih Doctoral Program, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia; Department of Histology, Faculty of Medicine, Universitas Islam Indonesia, Yogyakarta, Indonesia
  • Teguh Aryandono Department of Surgery, Oncology Division, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia
  • Sitarina Widyarini Department of Pathology, Faculty of Veterinary Medicine, Universitas Gadjah Mada, Yogyakarta, Indonesia
  • Sri Herwiyanti Department of Histology, Faculty of Medicine, Public Health and Nursing, Universitas Gadjah Mada, Yogyakarta, Indonesia



7,12-Dimethylbenzanthracene, Estrogen receptor, Progesterone receptor, Human epidermal growth factor receptor 2, Ki67


BACKGROUND: Animal models with mammary cancer that closely mimic human breast cancer for treatment development purposes are still required. Induction of 7,12-dimethylbenzanthracene (DMBA) to rats shows the histopathological features and mammary cancer characterization similar to humans. Examinations of estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), and Ki67 expressions are crucial in deciding the treatment and prognosis of breast cancer.

AIM: This research aimed to view histopathology images of mammary glands and expressions of ER, PR, Ki67, and HER2 of DMBA-induced rats.

METHODS: After 1-week adaptation, 11 5-weeks-old female rats were induced with 20 mg/kg body weight (BW) of DMBA 2 times a week for 5 weeks. On week 29, nodules taken from the mammary gland were examined for hematoxylin-eosin staining and immunohistochemistry with p63, ER, PR, HER2, and Ki67 antibodies. The grading score used the Nottingham Grading System and molecular classifications based on St. Gallen 2013.

RESULTS: Six rats had nodules, but the histopathologic features of one nodule showed normal mammary gland without cancer. The histopathological type of mammary cancer was cribriform carcinoma, comedo carcinoma, lipid-rich carcinoma, adenocarcinoma squamous, and adenomyepithelioma. Histopathological grading showed 60% of grade 3 and 40% of grade 2. P63 expression showed 60% positive and 40% negative. The frequency of ER, PR, HER2, and Ki67 of five nodules showed positivity: 40%, 60%, 60%, and 60%, respectively. Molecular subtypes of Luminal A, B, HER2, and triple-negative were 0%, 60%, 20%, and 20%, respectively.

CONCLUSION: Histopathological features and molecular subtype of mammary cancer on rats induced with 20 mg/kg BW of DMBA showed similarity to human breast cancer.


Download data is not yet available.


Metrics Loading ...

Plum Analytics Artifact Widget Block


World Health Organization. GLOBOCAN. 360 Indonesia Fact Sheet. Geneva: World Health Organization; 2020. Available from: [Last accessed on 2021 May 10].

Carioli G, Bertuccio P, Boffetta P, Levi F, La Vecchia C, Negri E, et al. European cancer mortality predictions for the year 2020 with a focus on prostate cancer. Ann Oncol. 2020;31(5):650-8. PMid:32321669 DOI:

Costa E, Ferreira-gonçalves T, Chasqueira G, Cabrita S. Experimental models as refined translational tools for breast cancer research. Sci Pharm. 2020;88(32):1-29. DOI:

Liu Y, Yin T, Feng Y, Cona MM, Huang G, Liu J, et al. Mammalian models of chemically induced primary malignancies exploitable for imaging-based preclinical theragnostic research. Quant Imaging Med Surg. 2015;5(5):708-29. PMid:26682141

Dai X, Li T, Bai Z, Yang Y, Liu X, Zhan J, et al. Breast cancer intrinsic subtype classification, clinical use and future trends. Am J Cancer Res. 2015;5(10):2929-43. PMid:26693050 DOI:

Nikolov S, Enchev E, Minkov G, Dimitrov E, Ivanova K, Gulubova M, et al. The role of the molecular subtypes in the prognosis of cancer patients. Open Access Maced J Med Sci. 2020;8(B):133-8. DOI:

Cheung SY, Yuen MT, Choi HL, Cheng HK, Huang Y, Chen S, et al. An expression study of hormone receptors in spontaneously developed, carcinogen-induced and hormone-induced mammary tumors in female Noble rats. Int J Oncol. 2003;22(6):1383-95. PMid:12739009 DOI:

Russo J. Significance of rodent mammary tumors for human risk assessment. Toxicol Pathol. 2015;43(2):145. PMid:25714400 DOI:

Charan J, Biswas T. How to calculate sample size for different study designs in medical research? Indian J Psychol Med. 2013;5(2):121-6. PMid:24049221 DOI:

Goldschmidt MH, Peña L, Rasotto R, Zappulli V. Classification and grading of canine mammary tumors. Vet Pathol. 2011;48(1):117-131. PMid:21266722 DOI:

Rudmann D, Cardiff R, Chouinard L, Goodman D, Küttler K, Marxfeld H, et al. Proliferative and nonproliferative lesions of the rat and mouse mammary, Zymbal’s, preputial, and clitoral glands. Toxicol Pathol. 2012;40 (6 Suppl):7-39. PMid:22949413 DOI:

Nascimento RG do, Otoni KM. Histological and molecular classification of breast cancer: What do we know? Mastology. 2020;30:e20200024. DOI:

Elston C, Ellis I. Pathological prognostic factors in breast cancer. I. The value of histological grade in breast cancer: Experience from a large study with long-term follow-up. Histopathology. 1991;19(5):403-10. PMid:25246403 DOI:

Goldhirsch A, Winer EP, Coates AS, Gelber RD, Thürlimann B, Panel HS. Personalizing the treatment of women with early breast cancer: Highlights of the St gallen international expert consensus on the primary therapy of early breast cancer 2013. Ann Oncol. 2013;24(9):2206-23. PMid:23917950 DOI:

Aiad H, Samaka R, Asaad N, Kandil M, Shehata M, Miligy I. Relationship of CK8/18 expression pattern to breast cancer immunohistochemical subtyping in Egyptian patients. ecancer Med Sci. 2014;8:404. PMid:24605136 DOI:

Reisenbichler ES, Balmer NN, Adams AL, Pfeifer JD, Hameed O. Luminal cytokeratin expression profiles of breast papillomas and papillary carcinomas and the utility of a cytokeratin 5/p63/ cytokeratin 8/18 antibody cocktail in their distinction. Mod Pathol. 2011;24(2):185-93. PMid:21076459 DOI:

Williams DL. Ocular disease in rats: A review. Vet Ophthalmol. 2002;5(3):183-91. PMid:12236869 DOI:

Smith JB, Mangkoewidjojo S. Pemeliharaan Pembiakan dan Penggunaan Hewan Percobaan di Daerah Tropis. Jakarta: Penerbit Universitas Indonesia; 1988.

Pritchett-Corning KR, Cosentino J, Clifford CB. Contemporary prevalence of infectious agents in laboratory mice and rats. Lab Anim. 2009;43(2):165-73. PMid:19015179 DOI:

McInnes EF, Rasmussen L, Fung P, Auld AM, Alvarez L, Lawrence DA, et al. Prevalence of viral, bacterial and parasitological diseases in rats and mice used in research environments in Australasia over a 5-y period. Lab Anim (NY). 2011;40(11):341-50. PMid:22012194 DOI:

Jangir BL, Chavhan S, Kurkure N, Chopade N. Diseases of laboratory rats. Livest Line. 2010;4(2):10-12.

Arora R, Bhushan S, Kumar R, Mannan R, Kaur P, Singh AP, et al. Hepatic dysfunction induced by 7, 12-dimethylbenz(α)anthracene and its obviation with erucin using enzymatic and histological changes as indicators. PLoS One. 2014;9(11):e112614. PMid:25390337 DOI:

Al-Asady AM, Ghaleb NK, Alnasrawi AM, Alhamed TA. Effect of carcinogenic substance (7,12 dimethylbenz [a] anthracene (dmba)) on tissue, hematology character and enzyme activity in rat. Indian J Forensic Med Toxicol. 2020;14(1):1172-76.

Gasparoto TH, De Oliveira CE, De Freitas LT, Pinheiro CR, Hori JI, Garlet GP, et al. Inflammasome activation is critical to the protective immune response during chemically induced squamous cell carcinoma. PLoS One. 2014;9(9):e107170. PMid:25268644 DOI:

Miyata M, Furukawa M, Takahashi K, Gonzalez FJ, Yamazoe Y. Mechanism of 7,12-dimethylbenz[a]anthracene-induced immunotoxicity: role of metabolic activation at the target organ. Jpn J Pharmacol. 2001;86(3):302-9. PMid:11488430 DOI:

Kocdor H, Cehreli R, Kocdor A, Sis B, Yilmaz O, Canda T, et al. Effects of selenium in Wistar rats. J Toxicol Environ Health. 2007;68(9):693-701. DOI:

Alvarado A, Lopes AC, Faustino-Rocha AI, Cabrita MS, Ferreira R, Oliveira PA, et al. Animal and in vitro models prognostic factors in MNU and DMBA-induced mammary tumors in female rats. Pathol Pract. 2017;213:441-6. DOI:

Russo J, Gusterson BA, Rogers AE, Russo IH, Wellings SR, Van Zwieten MJ. Comparative study of human and rat mammary tumorigenesis. Lab Investig. 1990;62(3):217-51. DOI:

Wahyuniari IA, Arijana IG, Sriwidyani NP, Wiryanthini IA, Suwito H, Widyarini S, et al. The anticancer activity of (e)-1-(4’- aminophenyl)-3-phenylprop-2-en-1-on against DMBA-induced mammary cancer in Sprague Dawley rat through the regulation of microRNA-21 expression. Bali Med J. 2017;6(3):589. DOI:

Guan B, Wang H, Cao S, Rao Q, Wang Y, Zhu Y, et al. Lipid-rich carcinoma of the breast clinicopathologic analysis of 17 cases. Ann Diagn Pathol. 2011;15(4):225-32. PMid:21396871 DOI:

Rehm S. Chemical induced mammary gland adenomyoepitheliomas and myoepithelial carcinomas of mice: Immunohistochemical and ultrastructural features. Am J Pathol. 1990;136(3):575-84. PMid:1690510

Machida Y, Imai T. Different properties of mammary carcinogenesis induced by two chemical carcinogens, DMBA and PhIP, in heterozygous BALB/c Trp53 knockout mice. Oncol Lett. 2021;22(4):738. PMid:34466150 DOI:

Merrill ML, Harper R, Birnbaum LS, Cardiff RD, Threadgill DW. Maternal dioxin exposure combined with a diet high in fat increases mammary cancer incidence in mice. Environ Health Perspect. 2010;118(5):596. PMid:20435547 DOI:

Intagliata E, Gangi S, Trovato C, Vecchio R, Strazzanti A. Benign adenomyoepitelioma of the breast: Presentation of two rare cases and review of literature. Int J Surg Case Rep. 2020;67:1-4. PMid:31991375 DOI:

O’Neil M, Fan F, Damjanov I. Adenomyoepithelioma of the preast. Lab Med. 2008;39(8):477-80. PMid:23627458 DOI:

Cheng Z, Han T, Zhang X, Li X, Li H, Gu J. Prognostic factors for breast cancer squamous cell carcinoma and nomogram development for prediction: Population-based research. Transl Cancer Res. 2019;8(5):2014-23. DOI:

Costa I, Solanas M, Escrich E. Histopathologic characterization of mammary neoplastic lesions induced with 7,12 dimethylbenz(alpha) anthracene in the rat: A comparative analysis with human breast tumors. Arch Pathol Lab Med. 2002;126(8):915-27. PMid:12171489 DOI:

Carraro DM, Elias EV, Andrade VP. Ductal carcinoma in situ of the breast: Morphological and molecular features implicated in progression. Biosci Rep. 2014;34(1):19-28. PMid:27919043 DOI:

Lopes AC, Cova TF, Pais AA, Pereira JL, Colaç B, Cabrita AM. Improving discrimination in the grading of rat mammary tumors using two-dimensional mapping of histopathological observations. Exp Toxicol Pathol. 2014;66(1):73-80. PMid:24168877 DOI:

Atif N. Role of immunohistochemical markers in breast cancer and their correlation with grade of tumour, our experience. Int Clin Pathol J. 2020;6(3):141-5. DOI:

Furqan M, Pohan PU. Relationship of histopathology grading with molecular subtypes of breast cancer patients in Haji Adam Malik General Hospital 2016-2018. Scr SCORE. Sci Med J. 2020;2(1):28-37. DOI:

Rakha EA, El-Sayed ME, Lee AH, Elston CW, Grainge MJ, Hodi Z. Prognostic significance of Nottingham histologic grade in invasive breast carcinoma. J Clin Oncol. 2008;26(19):3153-8. PMid:18490649 DOI:

Cheun JH, Jung J, Lee ES, Rhu J, Lee HB, Lee KH, et al. Intensity of metastasis screening and survival outcomes in patients with breast cancer. Sci Rep. 2021;11(1):2851. PMid:33531549 DOI:

Narisuari ID, Manuaba IB. Prevalence and characteristics of breast cancer patients at the surgical oncology polyclinic of Sanglah Hospital, Bali, Indonesia in 2016. Int Sains Med. 2020;11(1):183-9. DOI:

Feng M, Feng C, Yu Z, Fu Q, Ma Z, Wang F, et al. Histopathological alterations during breast carcinogenesis in a rat model induced by estrogen-progestogen combinations. Int J Clin Exp Med. 2015;8(1):346-57. PMid:25785005

Gallo D, Ferrandina G, Giacomelli S, Fruscella E, Zannoni GF, Morazzoni P, et al. Dietary soy modulation of biochemical parameters in DMBA-induced mammary tumors. Cancer Lett. 2002;186(1):43-8. PMid:12183074 DOI:

Fragomeni SM, Sciallis A, Jeruss JS. Molecular subtypes and local-regional control of breast cancer Simona. Surg Oncol Clin North Am. 2018;27(1):95-120. PMid:29132568 DOI:

Zhao JA, Chen JJ, Ju YC, Wu JH, Geng CZ, Yang HC. The effect of childbirth on carcinogenesis of DMBA-induced breast cancer in female SD rats. Chin J Cancer. 2011;30(11):779-85. PMid:22035859 DOI:

Varga Z, Noske A, Ramach C, Padberg B, Moch H. Assessment of HER2 status in breast cancer: Overall positivity rate and accuracy by fluorescence in situ hybridization and immunohistochemistry in a single institution over 12 years: A quality control study. BMC Cancer. 2013;13:615. PMid:24377754 DOI:

Sheikhpour E, Taghipour S. The differences of age, tumor grade, and Her2 amplification in Estrogen and progesterone receptor status in patients with breast cancer. Int J Cancer Manag. 2018;11(8):8-11. DOI:

Nanto SS, Muhartono, Wulan AJ. Role of estrogen receptor (ER), progesterone receptor (PR), and human epidermal growth factor receptor 2 (HER-2) to predict breast cancer clinical stage. J Agromed Unila. 2017;4(2):256-9.

Ma Z, Kim YM, Howard EW, Feng X, Kosanke SD, Yang S, et al. DMBA promotes ErbB2mediated carcinogenesis via ErbB2 and estrogen receptor pathway activation and genomic instability. Oncol Rep. 2018;40(3):1632-40. PMid:30015966 DOI:

Arnetha TS, Hernowo BS, Adha MJ, Rezano A. Relationship between molecular subtypes and overall survival of breast cancer in Bandung. Biomed Pharmacol J. 2020;13(3):1543-48. DOI:

Kanyilmaz G, Yavuz BB, Aktan M, Karaagac M, Uyar M, Findik S. Prognostic importance of Ki-67 in breast cancer and its relationship with other prognostic factors. Eur J Breast Heal. 2019;15(4):256-61. PMid:31620685 DOI:

Kermani TA, Kermani IA, Faham Z, Dolatkhah R. Ki-67 status in patients with primary breast cancer and its relationship with other prognostic factors. Biomed Res Ther. 2019;6(2):2986-91. PMid:23674192 DOI:

Hashmi AA, Hashmi KA, Irfan M, Khan SM, Edhi MM, Ali JP, et al. Ki67 index in intrinsic breast cancer subtypes and its association with prognostic parameters. BMC Res. 2019;12(1):605. PMid:31547858 DOI:

Currier N, Solomon SE, Demicco EG, Chang DLF, Farago M, Ying H, et al. Oncogenic signaling pathways activated in DMBA-induced mouse mammary tumors. Toxicol Pathol. 2005;33(6):726-37. PMid:16263698 DOI:

Asri A, Mayorita P, Khambri D. Relationship of ki-67 expression with histopathological characteristics in triple negative breast cancer. Maj Kedokt Andalas. 2015;38(3):165-72. DOI:

Sen TA, Poe J, Yeong S, Peng C, Lai T, Hui C, et al. The role of Ki-67 in Asian triple negative breast cancers: A novel combinatory panel approach. Virchows Arch. 2019;475(6):709-25. PMid:31407032 DOI:

Wu Q, Ma G, Deng Y, Luo W, Zhao Y, Li W, et al. Prognostic value of ki-67 in patients with resected triple-negative breast cancer: A meta-analysis. Front Oncol Front. 2019;9:1068. PMid:31681601 DOI:




How to Cite

Fidianingsih I, Aryandono T, Widyarini S, Herwiyanti S. Profile of Histopathological Type and Molecular Subtypes of Mammary Cancer of DMBA-induced Rat and its Relevancy to Human Breast Cancer. Open Access Maced J Med Sci [Internet]. 2022 Jan. 18 [cited 2024 Jul. 23];10(A):71-8. Available from:

Most read articles by the same author(s)