Advanced Imaging Approaches to Characterize Stromal and Metabolic Changes in In Vivo Mammary Tumor Models
Technical Report,01 Mar 2012,28 Feb 2015
University of Wisconsin, Madison Madison United States
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My goal is to investigate the connection between cancer cell progression and the cellular microenvironment, specifically by examining the effects of collagen density on cellular metabolism in breast cancer cells. Normal MCF10A and invasive MCF10-Ca1d breast epithelia cells were cultured in 2D and analyzed using high resolution to examine intracellular effects from cellular stress. To stress cellular metabolism, the cells were treated with either 1 percent O2 or DFOM, to mimic a hypoxic response, or 2DG to induce a hypoglycemic environment. Fluorescence lifetime data were then collected for the NADPH and FAD within the cells. DFOM, 1 percent O2, and 2DG treatment caused a decrease in the free fraction of NADPH and in lactate production, indicating the hypoxia response in MCF10a and MCF10-Ca1d cells may not be as simple as an increase in glycolysis. Utilizing the mammary imaging window technique in GFP-CFMS transgenic mouse, tumor cells decrease in the free fraction of NADPH compared to other cells types in and around the tumor. We did not see significant changes in fluorescence lifetime of NADPH between the PyVTCol1a1 mice and PyVTwild-type mice.