title: Functional and molecular characterisation of EO771.LMB tumours, a new C57BL/6-mouse-derived model of spontaneously metastatic mammary cancer.
abstract: The translation of basic research into improved therapies for breast cancer patients requires relevant preclinical models that incorporate spontaneous metastasis. We have completed a functional and molecular characterisation of a new isogenic C57BL/6 mouse model of breast cancer metastasis, comparing and contrasting it with the established BALB/c 4T1 model. Metastatic EO771.LMB tumours were derived from poorly metastatic parental EO771 mammary tumours. Functional differences were evaluated using both in vitro assays and spontaneous metastasis assays in mice. Results were compared to non-metastatic 67NR and metastatic 4T1.2 tumours of the 4T1 model. Protein and transcript levels of markers of human breast cancer molecular subtypes were measured in the four tumour lines, as well as p53 (Tp53) tumour-suppressor gene status and responses to tamoxifen in vivo and in vitro. Array-based expression profiling of whole tumours identified genes and pathways that were deregulated in metastatic tumours. EO771.LMB cells metastasised spontaneously to lung in C57BL/6 mice and displayed increased invasive capacity compared with parental EO771. By immunohistochemical assessment, EO771 and EO771.LMB were basal-like, as was the 4T1.2 tumour, whereas 67NR had a luminal phenotype. Primary tumours from all lines were negative for progesterone receptor, Erb-b2/Neu and cytokeratin 5/6, but positive for epidermal growth factor receptor (EGFR). Only 67NR displayed nuclear estrogen receptor alpha (ERα) positivity. EO771 and EO771.LMB expressed mutant p53, whereas 67NR and 4T1.2 were p53-null. Integrated molecular analysis of both the EO771/EO771.LMB and 67NR/4T1.2 pairs indicated that upregulation of matrix metalloproteinase-3 (MMP-3), parathyroid hormone-like hormone (Pthlh) and S100 calcium binding protein A8 (S100a8) and downregulation of the thrombospondin receptor (Cd36) might be causally involved in metastatic dissemination of breast cancer.
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, VIC 3010, Australia Department of Pathology, University of Melbourne, Parkville, VIC 3010, Australia Department of Pharmacology & Therapeutics, University of Melbourne, Parkville, VIC 3010, Australia.
authors: Cameron N
authors: CN
authors: Johnstone
authors: Angiogenesis and Metastasis Research, Royal College of Surgeons in Ireland, 123 St Stephen's Green, Dublin 2, Ireland.
authors: Yvonne E
authors: YE
authors: Smith
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Yuan
authors: Y
authors: Cao
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Allan D
authors: AD
authors: Burrows
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Ryan S N
authors: RS
authors: Cross
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Xiawei
authors: X
authors: Ling
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Richard P
authors: RP
authors: Redvers
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Judy P
authors: JP
authors: Doherty
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia Morgan Welch Inflammatory Breast Cancer Research and Clinic, Department of Breast Medical Oncology, The University of Texas, MD Anderson Cancer Center, Houston, TX 77030, USA.
authors: Bedrich L
authors: BL
authors: Eckhardt
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Anthony L
authors: AL
authors: Natoli
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Christina M
authors: CM
authors: Restall
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Erin
authors: E
authors: Lucas
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Helen B
authors: HB
authors: Pearson
authors: Department of Anatomical Pathology, Royal Melbourne Hospital, Parkville, VIC 2010, Australia.
authors: Siddhartha
authors: S
authors: Deb
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, VIC 3010, Australia.
authors: Kara L
authors: KL
authors: Britt
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Alexandra
authors: A
authors: Rizzitelli
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia.
authors: Jason
authors: J
authors: Li
authors: Angiogenesis and Metastasis Research, Royal College of Surgeons in Ireland, 123 St Stephen's Green, Dublin 2, Ireland.
authors: Judith H
authors: JH
authors: Harmey
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, VIC 3010, Australia Department of Pathology, University of Melbourne, Parkville, VIC 3010, Australia normand.pouliot@petermac.org robin.anderson@petermac.org.
authors: Normand
authors: N
authors: Pouliot
authors: Research Division, Peter MacCallum Cancer Centre, St Andrew's Place, East Melbourne, VIC 3002, Australia Sir Peter MacCallum Department of Oncology, University of Melbourne, Parkville, VIC 3010, Australia Department of Pathology, University of Melbourne, Parkville, VIC 3010, Australia normand.pouliot@petermac.org robin.anderson@petermac.org.
authors: Robin L
authors: RL
authors: Anderson
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000378:metabolism; D045744:Cell Line, Tumor; D004195:Disease Models, Animal; D047628:Estrogen Receptor alpha / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D005260:Female; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007150:Immunohistochemistry; D015674:Mammary Neoplasms, Animal / Q000145:classification / Q000188:drug therapy / Q000235:genetics / Q000473:pathology*; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D009362:Neoplasm Metastasis / Q000235:genetics / Q000473:pathology*; D009363:Neoplasm Proteins / Q000378:metabolism; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013629:Tamoxifen / Q000494:pharmacology / Q000627:therapeutic use; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; D047628:Estrogen Receptor alpha; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; D013629:Tamoxifen
keywords: Breast cancer; Estrogen receptor alpha; Metastasis; Syngeneic preclinical models; Tumour subtyping
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