Estrogen-dependent expression and subcellular localization of the tight junction protein claudin-4 in HEC-1A endometrial cancer cells

Cuevas,Maria  E.; Gaska,Jenna  M.; Gist,Andrea  C.; King,Jonathan  M.; Sheller,Rebecca  A.; Todd,Maria  C.

doi:10.3892/ijo.2015.3030

Estrogen-dependent expression and subcellular localization of the tight junction protein claudin-4 in HEC-1A endometrial cancer cells

Authors:
- Maria E. Cuevas
- Jenna M. Gaska
- Andrea C. Gist
- Jonathan M. King
- Rebecca A. Sheller
- Maria C. Todd
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Affiliations: Biology Department, Southwestern University, Georgetown, TX 78626, USA, Biology Department, Trinity University, San Antonio, TX 78212, USA
Published online on: June 4, 2015 https://doi.org/10.3892/ijo.2015.3030
Pages: 650-656

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Abstract

Endometrial cancer is the most common female reproductive cancer in the United States and is associated with deregulated tight junction protein expression. Given the highly estrogen-responsive nature of this tissue, we investigated the effects of estrogen and its agonist, 4-OH TAM, on the expression and subcellular localization of the tight junction protein claudin-4 (CLDN-4), in HEC-1A endometrial cancer cells. In untreated HEC-1A cells, we observed dramatic overexpression of claudin-4 protein. In addition, differential detergent extraction analysis indicated that claudin-4 was localized primarily in the membrane but also found in the cytosolic, nuclear and cytoskeletal fractions. Upon exposure of HEC-1A to estradiol (E2), we observed a biphasic effect both on the overall expression of claudin-4 protein and on its cytosolic and cytoskeletal presence as demonstrated by immunoblot analysis. Immunofluorescence analysis also revealed a biphasic effect of E2 on claudin-4 expression. In contrast, we observed no changes in expression levels nor in the subcellular distribution patterns of claudin-4 in HEC-1A cells treated with different concentrations of 4-OH TAM. The intracellular presence of CLDN-4 coupled with the biphasic effects of E2 on CLDN-4 expression in the cytoskeleton suggest that this protein may be involved in cell signaling to and from TJs.

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