Gene expression in tumor cells and stroma in dsRed 4T1 tumors in eGFP-expressing mice with and without enhanced oxygenation

BMC Cancer

Research Article

Ingrid Moen¹  Per Ø Enger¹  Linda Sleire¹  Martha Chekenya¹  Jian Wang¹  Rolf K Reed¹  Linda EB Stuhr¹  Charlotte Jevne¹  Karl-Henning Kalland²  Anne M Øyan²  Lars A Akslen³ 

[1] Department of Biomedicine, University of Bergen, Jonas Lies vei 91, 5009, Bergen, Norway;The Gade Institute, University of Bergen, Bergen, Norway;Department of Microbiology, Haukeland University Hospital, Bergen, Norway;The Gade Institute, University of Bergen, Bergen, Norway;Department of Pathology, Haukeland University Hospital, Bergen, Norway;

关键词: Stromal Cell;    Mammary Tumor;    Nikon Corporation;    Mammary Tumor Model;    eGFP Mouse;   

DOI  :  10.1186/1471-2407-12-21

 received in 2011-12-14, accepted in 2012-01-17,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundThe tumor microenvironment is pivotal in tumor progression. Thus, we aimed to develop a mammary tumor model to elucidate molecular characteristics in the stroma versus the tumor cell compartment by global gene expression. Secondly, since tumor hypoxia influences several aspects of tumor pathophysiology, we hypothesized that hyperoxia might have an inhibitory effect on tumor growth per se. Finally, we aimed to identify differences in gene expression and key molecular mechanisms, both in the native state and following treatment.Methods4T1 dsRed breast cancer cells were injected into eGFP expressing NOD/SCID mice. Group 1 was exposed to 3 intermittent HBO treatments (Day 1, 4 and 7), Group 2 to 7 daily HBO treatments (both 2.5bar, 100% O2, à 90 min), whereas the controls were exposed to a normal atmosphere. Tumor growth, histology, vascularisation, cell proliferation, cell death and metastasis were assessed. Fluorescence-activated cell sorting was used to separate tumor cells from stromal cells prior to gene expression analysis.ResultsThe purity of sorted cells was verified by fluorescence microscopy. Gene expression profiling demonstrated that highly expressed genes in the untreated tumor stroma included constituents of the extracellular matrix and matrix metalloproteinases. Tumor growth was significantly inhibited by HBO, and the MAPK pathway was found to be significantly reduced. Immunohistochemistry indicated a significantly reduced microvessel density after intermittent HBO, whereas daily HBO did not show a similar effect. The anti-angiogenic response was reflected in the expression trends of angiogenic factors.ConclusionsThe present in vivo mammary tumor model enabled us to separate tumor and stromal cells, and demonstrated that the two compartments are characterized by distinct gene expressions, both in the native state and following HBO treatments. Furthermore, hyperoxia induced a significant tumor growth-inhibitory effect, with significant down-regulation of the MAPK pathway. An anti-angiogenic effect after intermittent HBO was observed, and reflected in the gene expression profile.

【 授权许可】

CC BY   
© Moen et al; licensee BioMed Central Ltd. 2012