Estrogen and tamoxifen up-regulate FXYD3 on breast cancer cells: assessing the differential roles of ER α and ZEB1.

Increased expression of the FXYD3 family of proteins has been associated with lung, colorectal, bladder and pancreatic cancers, and recent evidence suggests that elevated FXYD3 may promote tumor cell proliferation in breast cancer as well. However, factors involved in up-regulating the expression of FXYD3 in breast cancer have not been identified. We evaluated whether estrogen and the selective estrogen receptor modulator tamoxifen could regulate the expression of FXYD3 on breast cancer cells. Estrogen receptor (ER) α-positive MCF-7 and ER α-negative MDA-MB-231 human breast cancer cells used in our studies were treated with estrogen, tamoxifen or the combination of these agents. Relative expression of FXYD3 was assessed using fluorochrome-tagged antibodies and a fluorescence cytometer. We found that estrogen and tamoxifen, used alone or in combination, significantly increased FXYD3 on MCF-7 cells. FXYD3 levels did not increase compared to the control samples when ER α-negative 231 cells were treated with estrogen or tamoxifen, alone or in combination, indicating that ER α was required for the increased FXYD3 response. We showed that ER α associates with the transcription factor ZEB1 in MCF-7 cells, and that decreasing ZEB1 protein expression using siRNA disrupts the ability of estrogen, but not tamoxifen, to increase FXYD3 in MCF-7 cells. Our results indicate that there may be two mechanisms, both involving ER α and one requiring ZEB1, through which FXYD3 may be increased by estrogen and tamoxifen in breast cancer cells. Ongoing research endeavors are focusing on identifying cellular components through which estrogen and tamoxifen, alone or in combination, differentially regulate FXYD3 expression in human breast cancer cells.