Effect of FGF Overexpression in Immortalized Mammary Epithelial Cells.
Annual rept. 15 Jun 94-14 Jun 95,
GEORGETOWN UNIV WASHINGTON DC
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Using MCF7 cellsa human breast carcinoma cell line which is estrogen receptor positionour lab built a system in which transfection of either FClF4 or FGFi into this cell line allows cells to form progressively growing tumors in athymic nude mice without estrogen supplementation. The overexpression of FGFs also forms micrometastases in different organs. The hypothesis to be tested is that FGFs are also capable of increasing the malignant potential of breast epithelial cells that are not as far along on a progression pathway as MCF7 cancer cells. We chose MCFiOA cells, which is a spontaneously immortalized mammary epithelial cell line to study this hypothesis. The background screening of this cell line, at mRNA level, indicates that this cell line expresses high level of FGF2 , also expresses FGFRi , barely detectable FGFR2 -, and does not express FGF1, FGF4, and FGFR3. At the protein level, we demonstrated that a high amount of FGF-2 protein exists in MCF1OA cell lysate, but barely detectable levels are present in the conditioned media. We transfected MCF-lOA cells with FUF1 and FGF4. Using RNAse protection assay, we obtained transfected clones that express FGF4. Using Southern blot assay, the data shows the FUF4 and FUF1 genes are integrated into the MCF-10A genomic DNA.
- Anatomy and Physiology
- Medicine and Medical Research