Dipak Panigrahy, Matthew L. Edin, Craig R. Lee, Sui Huang, Diane R. Bielenberg, Catherine E. Butterfield, Carmen M. Barnés, Akiko Mammoto, Tadanori Mammoto, Ayala Luria, Ofra Benny, Deviney M. Chaponis, Andrew C. Dudley, Emily R. Greene, Jo-Anne Vergilio, Giorgio Pietramaggiori, Sandra S. Scherer-Pietramaggiori, Sarah M. Short, Meetu Seth, Fred B. Lih, Kenneth B. Tomer, Jun Yang, Reto A. Schwendener, Bruce D. Hammock, John R. Falck, Vijaya L. Manthati, Donald E. Ingber, Arja Kaipainen, Patricia A. D’Amore, Mark W. Kieran, Darryl C. Zeldin
Pharmacological modulation of EET levels controls primary tumor growth and metastasis.
(A) Systemic administration of an sEH inhibitor (tAUCB) stimulates primary LLC-GFP tumor growth. Images show representative tumors after 13 days of treatment. n = 6 mice/group; *P = 0.007. Scale bar: 1 cm. (B) Immunofluorescence double staining for VEGF and GFP (tumor cells) shows increased tumor cell expression of VEGF in LLC-GFP tumor cells from tAUCB- versus vehicle-treated mice. Green, GFP-stained tumor cells; red, VEGF-containing cells. Colocalization of red and green fluorescence (yellow) indicates tumor cells expressing VEGF (arrows). tAUCB-treated tumors have an increase in MECA-32–positive ECs (green). Scale bars: 20 μm. (C) Systemic administration of tAUCB and TUPS (10 mg/kg/d each) increases spontaneous B16F10 axillary lymph node metastasis 21 days after B16F10 resection. Representative axillary lymph nodes after 21 days of treatment are shown. Scale bars: 1 cm. n = 6 mice/group; *P = 0.029 versus control. (D) The EET antagonist 14,15-EEZE (0.21 mg/mouse) inhibits primary LLC growth (left panel), prolongs survival (middle panel), and reduces plasma VEGF levels (right panel) in a spontaneous LLC lung metastasis model. n = 5 mice/group; *P = 0.017, **P = 0.035, ***P = 0.044. (E) The EET antagonist 14,15-EEZE-mSI (0.21 mg/mouse) inhibits 14,15-EET–induced (15 μg/kg/d) spontaneous LLC metastasis. The stable EET metabolite 14,15-DHET (15 μg/kg/d) does not stimulate metastasis. Representative photographs on day 12 after LLC resection are shown. Scale bars: 1 cm. n = 5 mice/group; *P < 0.001, **P = 0.003 versus 14,15-EET.