In vivo bioluminescence imaging of tumor hypoxia dynamics of breast cancer brain metastasis in a mouse model.

Debabrata Saha, Henry Dunn, Heling Zhou, Hiroshi Harada, Masahiro Hiraoka, Ralph P. Mason, Dawen Zhao

Research output: Contribution to journalArticle

Abstract

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There is little knowledge about in situ, in vivo, tumor hypoxia during intracranial development of malignant brain tumors because of lack of efficient means to monitor it in these deep-seated orthotopic tumors. Bioluminescence imaging (BLI), based on the detection of light emitted by living cells expressing a luciferase gene, has been rapidly adopted for cancer research, in particular, to evaluate tumor growth or tumor size changes in response to treatment in preclinical animal studies. Moreover, by expressing a reporter gene under the control of a promoter sequence, the specific gene expression can be monitored non-invasively by BLI. Under hypoxic stress, signaling responses are mediated mainly via the hypoxia inducible factor-1α (HIF-1α) to drive transcription of various genes. Therefore, we have used a HIF-1α reporter construct, 5HRE-ODD-luc, stably transfected into human breast cancer MDA-MB231 cells (MDA-MB231/5HRE-ODD-luc). In vitro HIF-1α bioluminescence assay is performed by incubating the transfected cells in a hypoxic chamber (0.1% O2) for 24 hr before BLI, while the cells in normoxia (21% O2) serve as a control. Significantly higher photon flux observed for the cells under hypoxia suggests an increased HIF-1α binding to its promoter (HRE elements), as compared to those in normoxia. Cells are injected directly into the mouse brain to establish a breast cancer brain metastasis model. In vivo bioluminescence imaging of tumor hypoxia dynamics is initiated 2 wks after implantation and repeated once a week. BLI reveals increasing light signals from the brain as the tumor progresses, indicating increased intracranial tumor hypoxia. Histological and immunohistochemical studies are used to confirm the in vivo imaging results. Here, we will introduce approaches of in vitro HIF-1α bioluminescence assay, surgical establishment of a breast cancer brain metastasis in a nude mouse and application of in vivo bioluminescence imaging to monitor intracranial tumor hypoxia.

Original languageEnglish (US)
JournalJournal of visualized experiments : JoVE
Issue number56
StatePublished - 2011

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Bioluminescence
Hypoxia-Inducible Factor 1
Brain Neoplasms
Tumors
Brain
Breast Neoplasms
Neoplasm Metastasis
Imaging techniques
Neoplasms
Light
Cell Hypoxia
Genes
Luciferases
Reporter Genes
Photons
Nude Mice
Assays
Brain models
Tumor Hypoxia
Gene Expression

ASJC Scopus subject areas

  • Medicine(all)

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In vivo bioluminescence imaging of tumor hypoxia dynamics of breast cancer brain metastasis in a mouse model. / Saha, Debabrata; Dunn, Henry; Zhou, Heling; Harada, Hiroshi; Hiraoka, Masahiro; Mason, Ralph P.; Zhao, Dawen.

In: Journal of visualized experiments : JoVE, No. 56, 2011.

Research output: Contribution to journalArticle

Saha, Debabrata ; Dunn, Henry ; Zhou, Heling ; Harada, Hiroshi ; Hiraoka, Masahiro ; Mason, Ralph P. ; Zhao, Dawen. / In vivo bioluminescence imaging of tumor hypoxia dynamics of breast cancer brain metastasis in a mouse model. In: Journal of visualized experiments : JoVE. 2011 ; No. 56.
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