Poster Presentation 25th Lorne Cancer Conference 2013

The inflammation-resolving trihydroxy-docosahexaenoic acid derivative, resolvin D2 supports MCF-7 cell proliferation via activation of estrogen receptor (#106)

Nuha Alzaubai 1 , Michael Schuliga 1 , Trudi Harris 1 , John Li 2 , May Leong 2 , Mark Rizzacasa 2 , Alastair Stewart 1
  1. University of Melbourne, Melbourne, VIC, Australia
  2. school of chemistry, university of melbourne, melbourne , victoria, australia

Introduction: Resolvin D2(RvD2) is an inflammation-resolving tri-hydroxy lipid mediator generated endogenously from the omega-3 polyunsaturated fatty acid, docosahexaenoic acid (DHA)2. Formyl peptide receptors(FPRs) are G-protein coupled receptors. FPRs can bind a wide range of structurally diverse ligands, including proteins, small peptides and bioactive lipid mediators. They are expressed in both estrogen receptor-positive (MCF-7) and estrogen receptor-negative (MDA-MB-231) breast cancer cell lines. Some of the endogenous inflammation-resolving FPR ligands (annexin A1 and its N-terminal peptide Ac2-26, and the tri-hydroxy lipid mediator lipoxin A4) stimulate the proliferation of both MCF-7 and MDA-MB-231 cells through activation of FPRs1.
Aim: To investigate the impact of RvD2 on breast tumour cell proliferation.
Methods: Resolvin D2 was obtained by total chemical synthesis. Both MCF-7 and MDA-MB-231 were seeded in 24-well plates in 10% FCS media at 50,000 cells/well for 24 hours. Cells were then incubated with serum free media for another 24 hours before treatment with FCS 5% (v/v) or RvD2 0.1-100 nM. After 48 hours viable cells were enumerated. MCF-7 cells were transfected with estrogen response element (ERE)-controlled secretory alkaline phosphatase (SEAP) and pGL3 luciferase vector to account for transfection efficiencies. ERE activity was measured using a SEAP detection kit. MCF-7 cytosol was incubated with increasing concentrations of estradiol or RvD2 in the presence of 3H-E2.
Results: RvD2 stimulated the proliferation of MCF-7 cells, but not MDA-MB-231 cells. The proliferative effect of 100nM RvD2 was prevented by incubation with estrogen receptor antagonist, ICI 182,780 (100nM). RvD2 stimulated ERE activity but did not affect 3H-E2 binding to MCF-7 cytosol .
Conclusion: The data suggest that RvD2 induces the growth of MCF-7, but not MDA-MB-231 cells. RvD2 actions appear to be dependent on estrogen receptor but do not involve binding to the same site as estrogen.

  1. Khau, T., Langenbach, S. Y., Schuliga, M., Harris, T., Johnstone, C. N., Anderson, R. L., and Stewart, A. G. (2011). Annexin-1 signals mitogen-stimulated breast tumor cell proliferation by activation of the formyl peptide receptors (FPRs) 1 and 2. FASEB J 25, 483-496.
  2. Serhan, C. N., Krishnamoorthy, S., Recchiuti, A., and Chiang, N. (2011). Novel anti-inflammatory--pro-resolving mediators and their receptors. Curr Top Med Chem 11, 629-647.