Cell Fusion and Breast Cancer Metastasis
Final rept. 15 Aug 2007-14 Aug 2008
IOWA UNIV IOWA CITY
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This proposal had two overall objectives 1 Determine the frequency of breast cancer cell X host cell fusion events in experimental tumors and 2 Measure the metastatic capacity of cells derived from fusion events. We engineered murine mammary carcinoma cells with a reporter construct in which fluorescent protein mCherry expression was dependent upon Cre-mediated recombination. We demonstrated that delivery of Cre to reporter cells in vitro activated fluorescent protein expression demonstrating the utility of our system. We then injected cells orthotopically into the mammary fat pads of syngeneic hosts ubiquitously expressing Cre recombinase under control of the CMV promoter. We analyzed the resulting tumors for mCherry expression by FACS and histology. We did not detect any mCherry expressing cells by FACS nor did we see mCherry positive cells in tissue sections. We established that the FACS assay was sensitive enough to detect 1 in 1000 mCherry positive cells. Thus we were not able to identify cell fusion events in vivo using this approach.
- Anatomy and Physiology
- Medicine and Medical Research