Author:
Woock Alicia E.,Grible Jacqueline M.,Olex Amy L.,Harrell J. Chuck,Zot Patricija,Idowu Michael,Clevenger Charles V.
Abstract
AbstractIn breast cancer, prolactin-induced activation of the transcription factor STAT5a results from the phosphorylation of STAT5a tyrosine residue 694. However, its role in mammary oncogenesis remains an unsettled debate as STAT5a exhibits functional dichotomy with both pro-differentiative and pro-proliferative target genes. Phosphorylation of STAT5a serine residues, S726 and S780, may regulate STAT5a in such a way to underlie this duality. Given hematopoiesis studies showing phospho-serine STAT5a as necessary for transformation, we hypothesized that serine phosphorylation regulates STAT5a activity to contribute to its role in mammary oncogenesis, specifically in luminal breast cancer. Here, phosphorylation of S726-, S780-, and Y694-STAT5a in response to prolactin in MCF7 luminal breast cancer cells was investigated with STAT5a knockdown and rescue with Y694F-, S726A-, or S780A-STAT5a, where the phospho-sites were mutated. RNA-sequencing and subsequent Ingenuity Pathway Analysis predicted that loss of each phospho-site differentially affected both prolactin-induced gene expression as well as functional pathways of breast cancer (e.g. cell survival, proliferation, and colony formation). In vitro studies of anchorage-independent growth and proliferation confirmed distinct phenotypes: whereas S780A-STAT5a decreased clonogenicity, S726A-STAT5a decreased proliferation in response to prolactin compared to wild type STAT5a. Collectively, these studies provide novel insights into STAT5a activation in breast cancer pathogenesis.
Funder
National Center for Advancing Translational Sciences
Commonwealth Health Research Board
Publisher
Springer Science and Business Media LLC
Reference73 articles.
1. Ormandy, C. J. et al. Null mutation of the prolactin receptor gene produces multiple reproductive defects in the mouse. Genes Dev. 11(2), 167–178. https://doi.org/10.1101/gad.11.2.167 (1997).
2. Clevenger, C. V. et al. Expression of prolactin and prolactin receptor in human breast carcinoma. Evidence for an autocrine/paracrine loop. Am. J. Pathol. 146(3), 695–705 (1995).
3. Horseman, N. D. et al. Defective mammopoiesis, but normal hematopoiesis, in mice with a targeted disruption of the prolactin gene. EMBO J. 16(23), 6926–6935. https://doi.org/10.1093/emboj/16.23.6926 (1997).
4. Clevenger, C. V. & Plank, T. L. Prolactin as an autocrine/paracrine factor in breast tissue. J Mammary Gland Biol Neoplasia. 2(1), 59–68. https://doi.org/10.1023/a:1026325630359 (1997).
5. Ormandy, C. J. & Sutherland, R. L. Mechanisms of prolactin receptor regulation in mammary gland. Mol. Cell Endocrinol. 91(1), C1–C6. https://doi.org/10.1016/0303-7207(93)90247-H (1993).
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