論文

査読有り
2011年10月

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
  • Debabrata Saha
  • ,
  • Henry Dunn
  • ,
  • Heling Zhou
  • ,
  • Hiroshi Harada
  • ,
  • Masahiro Hiraoka
  • ,
  • Ralph P. Mason
  • ,
  • Dawen Zhao

56
記述言語
英語
掲載種別
研究論文(学術雑誌)
DOI
10.3791/3175
出版者・発行元
JOURNAL OF VISUALIZED EXPERIMENTS

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-1 alpha (HIF-1 alpha) to drive transcription of various genes. Therefore, we have used a HIF-1 alpha reporter construct, 5HRE-ODD-luc, stably transfected into human breast cancer MDA-MB231 cells (MDA-MB231/5HRE-ODD-luc). In vitro HIF-1 alpha bioluminescence assay is performed by incubating the transfected cells in a hypoxic chamber (0.1% O-2) for 24 hr before BLI, while the cells in normoxia (21% O-2) serve as a control. Significantly higher photon flux observed for the cells under hypoxia suggests an increased HIF-1 alpha 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-1 alpha 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.

リンク情報
DOI
https://doi.org/10.3791/3175
Web of Science
https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000209222400024&DestApp=WOS_CPL
ID情報
  • DOI : 10.3791/3175
  • ISSN : 1940-087X
  • Web of Science ID : WOS:000209222400024

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