Affiliation:
1. School of Life Science and Technology Tokyo Institute of Technology Yokohama Japan
2. Institute for Life and Medical Sciences Kyoto University Kyoto Japan
3. Center for Molecular Medicine Jichi Medical University Shimotsuke Japan
4. Graduate School of Informatics and Engineering The University of Electro‐Communications Chofu Japan
Abstract
AbstractTumors contain various stromal cells, such as immune cells, endothelial cells, and fibroblasts, which contribute to the development of a tumor‐specific microenvironment characterized by hypoxia and inflammation, and are associated with malignant progression. In this study, we investigated the activity of intratumoral hypoxia‐inducible factor (HIF), which functions as a master regulator of the cellular response to hypoxia and inflammation. We constructed the HIF activity‐monitoring reporter gene hypoxia‐response element‐Venus‐Akaluc (HVA) that expresses the green fluorescent protein Venus and modified firefly luciferase Akaluc in a HIF activity‐dependent manner, and created transgenic mice harboring HVA transgene (HVA‐Tg). In HVA‐Tg, HIF‐active cells can be visualized using AkaBLI, an ultra‐sensitive in vivo bioluminescence imaging technology that produces an intense near‐infrared light upon reaction of Akaluc with the D‐luciferin analog AkaLumine‐HCl. By orthotopic transplantation of E0771, a mouse triple negative breast cancer cell line without a reporter gene, into HVA‐Tg, we succeeded in noninvasively monitoring bioluminescence signals from HIF‐active stromal cells as early as 8 days after transplantation. The HIF‐active stromal cells initially clustered locally and then spread throughout the tumors with growth. Immunohistochemistry and flow cytometry analyses revealed that CD11b+F4/80+ macrophages were the predominant HIF‐active stromal cells in E0771 tumors. These results indicate that HVA‐Tg is a useful tool for spatiotemporal analysis of HIF‐active tumor stromal cells, facilitating investigation of the roles of HIF‐active tumor stromal cells in tumor growth and malignant progression.
Funder
Naito Foundation
Uehara Memorial Foundation
Subject
Cancer Research,Oncology,General Medicine
Cited by
1 articles.
订阅此论文施引文献
订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献