Establishment of a murine breast tumor model by subcutaneous or orthotopic implantation

Zhang,Yi; Zhang,Gan‑Lin; Sun,Xu; Cao,Ke‑Xin; Ma,Cong; Nan,Nan; Yang,Guo‑Wang; Yu,Ming‑Wei; Wang,Xiao‑Min

doi:10.3892/ol.2018.8113

Establishment of a murine breast tumor model by subcutaneous or orthotopic implantation

Authors:
- Yi Zhang
- Gan‑Lin Zhang
- Xu Sun
- Ke‑Xin Cao
- Cong Ma
- Nan Nan
- Guo‑Wang Yang
- Ming‑Wei Yu
- Xiao‑Min Wang
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Affiliations: Department of Oncology, Beijing Hospital of Traditional Chinese Medicine, Capital Medical University, Beijing 100010, P.R. China
Published online on: February 23, 2018 https://doi.org/10.3892/ol.2018.8113
Pages: 6233-6240
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

A number of murine models are used to mimic the pathology of breast cancer. Tissue inoculation and cell inoculation using orthotopic implantation (OS) and subcutaneous implantation (SQ) are commonly used to generate murine models to investigate cancer. However, limited information is available in regard to the variations of these methods. The present study compared growth, metastasis, survival and histopathology of tumors produced using OS and SQ to characterize features of the tumors produced by the two distinct methods. Additionally, the present study aimed at providing increased options for investigators when designing experiments. 4T1‑luc2 cell suspension or 4T1‑luc2 tissue suspension was inoculated using either OS or SQ into BALB/c mice. Tumor growth and metastasis were detected using an in vivo imaging system and calipers. Excised tumors and lung were assessed by tissue staining with hematoxylin and eosin, and the vessel marker cluster of differentiation 31. The results of the present study revealed that the cell suspension generated breast tumors of increased size, which was visualized and determined, following inoculation, using calipers at an earlier time point compared with tumors produced by tissue suspension. The increasing bioluminescent trend of OS tumors was more marked compared with that of SQ tumors. The volume of OS tumor was increased with decreased variation, compared with that of SQ tumors. In addition, the OS tumor exhibited increased microvessel density. Bioluminescent signals and histological results in regard to metastasis were consistent: OS implantation produced increased lung metastasis compared with that of SQ implantation, although they exhibited similar survival times. The results of the present study indicated that the inocula from distinct sources (tissue or cell) affected tumor growth. Furthermore, breast tumor progression and histopathological characteristics were distinct between OS and SQ, whereas OS exhibited increased malignant behavior. Understanding the characteristics of murine breast cancer models established by diverse methods may aid investigators to select appropriate animal models, according to the requirements of the study.

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1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Shi XJ, Au WW, Wu KS, Chen LX and Lin K: Mortality characteristics and prediction of female breast cancer in China from 1991 to 2011. Asian Pac J Cancer Prev. 15:2785–2791. 2014. View Article : Google Scholar : PubMed/NCBI
3	Light DW: Global drug discovery: Europe is ahead. Health Aff (Millwood). 28:w969–w977. 2009. View Article : Google Scholar : PubMed/NCBI
4	Ottewell PD, Coleman RE and Holen I: From genetic abnormality to metastases: Murine models of breast cancer and their use in the development of anticancer therapies. Breast Cancer Res Treat. 96:101–113. 2006. View Article : Google Scholar : PubMed/NCBI
5	Rashid OM and Takabe K: Animal models for exploring the pharmacokinetics of breast cancer therapies. Expert Opin Drug Metab Toxicol. 11:221–230. 2015. View Article : Google Scholar : PubMed/NCBI
6	Wagner KU: Models of breast cancer: Quo vadis, animal modeling? Breast Cancer Res. 6:31–38. 2004. View Article : Google Scholar : PubMed/NCBI
7	Donehower LA, Harvey M, Slagle BL, McArthur MJ, Montgomery CA Jr, Butel JS and Bradley A: Mice deficient for p53 are developmentally normal but susceptible to spontaneous tumours. Nature. 356:215–221. 1992. View Article : Google Scholar : PubMed/NCBI
8	Escobar Galvis ML, Jia J, Zhang X, Jastrebova N, Spillmann D, Gottfridsson E, van Kuppevelt TH, Zcharia E, Vlodavsky I, Lindahl U and Li JP: Transgenic or tumor-induced expression of heparanase upregulates sulfation of heparan sulfate. Nat Chem Biol. 3:773–778. 2007. View Article : Google Scholar : PubMed/NCBI
9	Matsui Y, Halter SA, Holt JT, Hogan BL and Coffey RJ: Development of mammary hyperplasia and neoplasia in MMTV-TGF alpha transgenic mice. Cell. 61:1147–1155. 1990. View Article : Google Scholar : PubMed/NCBI
10	Ursini-Siegel J, Schade B, Cardiff RD and Muller WJ: Insights from transgenic mouse models of ERBB2-induced breast cancer. Nat Rev Cancer. 7:389–397. 2007. View Article : Google Scholar : PubMed/NCBI
11	Menezes ME, Das SK, Emdad L, Windle JJ, Wang XY, Sarkar D and Fisher PB: Genetically engineered mice as experimental tools to dissect the critical events in breast cancer. Adv Cancer Res. 121:331–382. 2014. View Article : Google Scholar : PubMed/NCBI
12	Chakravarty G, Mathur A, Mallade P, Gerlach S, Willis J, Datta A, Srivastav S, Abdel-Mageed AB and Mondal D: Nelfinavir targets multiple drug resistance mechanisms to increase the efficacy of doxorubicin in MCF-7/Dox breast cancer cells. Biochimie. 124:53–64. 2016. View Article : Google Scholar : PubMed/NCBI
13	Li H, Pan GF, Jiang ZZ, Yang J, Sun LX and Zhang LY: Triptolide inhibits human breast cancer MCF-7 cell growth via downregulation of the ERα-mediated signaling pathway. Acta Pharmacol Sin. 36:606–613. 2015. View Article : Google Scholar : PubMed/NCBI
14	Habu S, Fukui H, Shimamura K, Kasai M, Nagai Y, Okumura K and Tamaoki N: In vivo effects of anti-asialo GM1. I. Reduction of NK activity and enhancement of transplanted tumor growth in nude mice. J Immunol. 127:34–38. 1981.PubMed/NCBI
15	Marangoni E, Vincent-Salomon A, Auger N, Degeorges A, Assayag F, de Cremoux P, de Plater L, Guyader C, De Pinieux G, Judde JG, et al: A new model of patient tumor-derived breast cancer xenografts for preclinical assays. Clin Cancer Res. 13:3989–3998. 2007. View Article : Google Scholar : PubMed/NCBI
16	Schuh JC: Trials, tribulations, and trends in tumor modeling in mice. Toxicol Pathol. 32 Suppl 1:S53–S66. 2004. View Article : Google Scholar
17	Talmadge JE, Singh RK, Fidler IJ and Raz A: Murine models to evaluate novel and conventional therapeutic strategies for cancer. Am J Pathol. 170:793–804. 2007. View Article : Google Scholar : PubMed/NCBI
18	Gravekamp C, Sypniewska R, Gauntt S, Tarango M, Price P and Reddick R: Behavior of metastatic and nonmetastatic breast tumors in old mice. Exp Biol Med (Maywood). 229:665–675. 2004. View Article : Google Scholar : PubMed/NCBI
19	Singh M, Ramos I, Asafu-Adjei D, Quispe-Tintaya W, Chandra D, Jahangir A, Zang X, Aggarwal BB and Gravekamp C: Curcumin improves the therapeutic efficacy of Listeria(at)-Mage-b vaccine in correlation with improved T-cell responses in blood of a triple-negative breast cancer model 4T1. Cancer Med. 2:571–582. 2013. View Article : Google Scholar : PubMed/NCBI
20	Takahashi K, Nagai N, Ogura K, Tsuneyama K, Saiki I, Irimura T and Hayakawa Y: Mammary tissue microenvironment determines T cell-dependent breast cancer-associated inflammation. Cancer Sci. 106:867–874. 2015. View Article : Google Scholar : PubMed/NCBI
21	Tao K, Fang M, Alroy J and Sahagian GG: Imagable 4T1 model for the study of late stage breast cancer. BMC Cancer. 8:2282008. View Article : Google Scholar : PubMed/NCBI
22	Zhou H, Roy S, Cochran E, Zouaoui R, Chu CL, Duffner J, Zhao G, Smith S, Galcheva-Gargova Z, Karlgren J, et al: M402, a novel heparan sulfate mimetic, targets multiple pathways implicated in tumor progression and metastasis. PLoS One. 6:e211062011. View Article : Google Scholar : PubMed/NCBI
23	Lee HS, Ha AW and Kim WK: Effect of resveratrol on the metastasis of 4T1 mouse breast cancer cells in vitro and in vivo. Nutr Res Pract. 6:294–300. 2012. View Article : Google Scholar : PubMed/NCBI
24	Mehta RR, Katta H, Kalra A, Patel R, Gupta A, Alimirah F, Murillo G, Peng X, Unni A, Muzzio M and Mehta RG: Efficacy and mechanism of action of Deguelin in suppressing metastasis of 4T1 cells. Clin Exp Metastasis. 30:855–866. 2013. View Article : Google Scholar : PubMed/NCBI
25	Gibson-D'Ambrosio RE, Samuel M and D'Ambrosio SM: A method for isolating large numbers of viable disaggregated cells from various human tissues for cell culture establishment. In Vitro Cell Dev Biol. 22:529–534. 1986. View Article : Google Scholar : PubMed/NCBI
26	Weigand A, Boos AM, Tasbihi K, Beier JP, Dalton PD, Schrauder M, Horch RE, Beckmann MW, Strissel PL and Strick R: Selective isolation and characterization of primary cells from normal breast and tumors reveal plasticity of adipose derived stem cells. Breast Cancer Res. 18:322016. View Article : Google Scholar : PubMed/NCBI
27	Weidner N, Semple JP, Welch WR and Folkman J: Tumor angiogenesis and metastasis-correlation in invasive breast carcinoma. N Engl J Med. 324:1–8. 1991. View Article : Google Scholar : PubMed/NCBI
28	Jordan VC: Proven value of translational research with appropriate animal models to advance breast cancer treatment and save lives: The tamoxifen tale. Br J Clin Pharmacol. 79:254–267. 2015. View Article : Google Scholar : PubMed/NCBI
29	Clarke R: Animal models of breast cancer: Their diversity and role in biomedical research. Breast Cancer Res Treat. 39:1–6. 1996. View Article : Google Scholar : PubMed/NCBI
30	Morioka CY, Saito S, Ohzawa K, Asano S, Hibino Y, Nakada Y, Kita KI and Watanabe A: Subcutaneously inoculated cells and implanted pancreatic cancer tissue show different patterns of metastases in Syrian golden hamsters. JOP. 1:183–190. 2000.PubMed/NCBI
31	Rao Q, You A, Guo Z, Zuo B, Gao X, Zhang T, Du Z, Wu C and Yin H: Intrahepatic tissue implantation represents a favorable approach for establishing orthotopic transplantation hepatocellular carcinoma mouse models. PLoS One. 11:e1482632016. View Article : Google Scholar
32	de la Cruz-Merino L, Barco-Sánchez A, Henao Carrasco F, Nogales Fernández E, Vallejo Benítez A, Brugal Molina J, Martínez Peinado A, Grueso López A, Ruiz Borrego M, Codes Manuel de Villena M, et al: New insights into the role of the immune microenvironment in breast carcinoma. Clin Dev Immunol. 2013:7853172013. View Article : Google Scholar : PubMed/NCBI
33	Rashid OM, Nagahashi M, Ramachandran S, Dumur C, Schaum J, Yamada A, Terracina KP, Milstien S, Spiegel S and Takabe K: An improved syngeneic orthotopic murine model of human breast cancer progression. Breast Cancer Res Treat. 147:501–512. 2014. View Article : Google Scholar : PubMed/NCBI
34	Rashid OM, Nagahashi M, Ramachandran S, Dumur CI, Schaum JC, Yamada A, Aoyagi T, Milstien S, Spiegel S and Takabe K: Is tail vein injection a relevant breast cancer lung metastasis model? J Thorac Dis. 5:385–392. 2013.PubMed/NCBI
35	Bibby MC: Orthotopic models of cancer for preclinical drug evaluation: Advantages and disadvantages. Eur J Cancer. 40:852–857. 2004. View Article : Google Scholar : PubMed/NCBI
36	Fidler IJ: The pathogenesis of cancer metastasis: The ‘seed and soil’ hypothesis revisited. Nat Rev Cancer. 3:453–458. 2003. View Article : Google Scholar : PubMed/NCBI
37	Nieman KM, Romero IL, Van Houten B and Lengyel E: Adipose tissue and adipocytes support tumorigenesis and metastasis. Biochim Biophys Acta. 1831:1533–1541. 2013. View Article : Google Scholar : PubMed/NCBI
38	Hammond E, Brandt R and Dredge K: PG545, a heparan sulfate mimetic, reduces heparanase expression in vivo, blocks spontaneous metastases and enhances overall survival in the 4T1 breast carcinoma model. PLoS One. 7:e521752012. View Article : Google Scholar : PubMed/NCBI
39	Kim JB, Urban K, Cochran E, Lee S, Ang A, Rice B, Bata A, Campbell K, Coffee R, Gorodinsky A, et al: Non-invasive detection of a small number of bioluminescent cancer cells in vivo. PLoS One. 5:e93642010. View Article : Google Scholar : PubMed/NCBI