Estrogen-mediated angiogenesis in thyroid tumor microenvironment is mediated through VEGF signaling pathways

Ameet Kamat, Shilpi Rajoria, Andrea George, Robert Suriano, Arul Kumaran Shanmugam, Uchechukwu Megwalu, Pradeep Bangalore Prakash, Raj Tiwari, Stimson Schantz

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Objectives: To explore the induction of a proangiogenic phenotype in endothelial cells in the thyroid tumor microenvironment by estrogen-treated thyroid cancer cells and to define the role of vascular endothelial growth factor (VEGF) in this interaction. Design: Cell-based in vitro systems analysis. Subjects: Thyroid tumor cell lines (BCPAP [papillary thyroid cancer] and ML-1 [follicular thyroid cancer]) were cultured with estradiol with and without an estrogen receptor (ER) inhibitor (fulvestrant or ICI) and used to treat human umbilical vein endothelial cells (HUVECs). Interventions: Immunofluorescence was used to confirm the presence of ERα and ERβ in BCPAP cells. Conditioned medium was then used to evaluate the induction of HUVEC tubulogenesis and migration. Secretion of VEGF in this medium was evaluated by Western blot analysis. The expression of phosphoinositide 3-kinase (PI3K), the initiator of a proangiogenic pathway, was evaluated with Western blot analysis of HUVEC lysates. The subsequent effects of an ER inhibitor (fulvestrant/ ICI) and a neutralizing VEGF antibody were also observed. Results: Estrogen receptor α and ERβ are expressed in thyroid cancer cells. Estrogen-stimulated ML-1 cells secreted an increased amount of VEGF likely as a result of ER signaling. In contact with this environment, HUVECs demonstrate enhanced tubulogenesis and migration. Western blot analysis documented estrogen-mediated upregulation of PI3K in HUVECs. These effects were mitigated by an ER inhibitor (fulvestrant/ICI) and a neutralizing VEGF antibody. Conclusions: Our data provide evidence that estrogen can induce a proangiogenic endothelial cell phenotype in the thyroid tumor microenvironment through ER and VEGF signaling. Our findings suggest that the effect of antiestrogenic therapy targeting tumor angiogenesis can be enhanced through VEGF inhibition.

Original languageEnglish (US)
Pages (from-to)1146-1153
Number of pages8
JournalArchives of Otolaryngology - Head and Neck Surgery
Volume137
Issue number11
DOIs
StatePublished - Nov 1 2011

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Tumor Microenvironment
Estrogen Receptors
Vascular Endothelial Growth Factor A
Thyroid Gland
Estrogens
Human Umbilical Vein Endothelial Cells
1-Phosphatidylinositol 4-Kinase
Western Blotting
Thyroid Neoplasms
Endothelial Cells
Phenotype
Antibodies
Conditioned Culture Medium
Systems Analysis
Tumor Cell Line
Cell Movement
Fluorescent Antibody Technique
Estradiol
Up-Regulation

ASJC Scopus subject areas

  • Otorhinolaryngology
  • Surgery

Cite this

Estrogen-mediated angiogenesis in thyroid tumor microenvironment is mediated through VEGF signaling pathways. / Kamat, Ameet; Rajoria, Shilpi; George, Andrea; Suriano, Robert; Shanmugam, Arul Kumaran; Megwalu, Uchechukwu; Prakash, Pradeep Bangalore; Tiwari, Raj; Schantz, Stimson.

In: Archives of Otolaryngology - Head and Neck Surgery, Vol. 137, No. 11, 01.11.2011, p. 1146-1153.

Research output: Contribution to journalArticle

Kamat, A, Rajoria, S, George, A, Suriano, R, Shanmugam, AK, Megwalu, U, Prakash, PB, Tiwari, R & Schantz, S 2011, 'Estrogen-mediated angiogenesis in thyroid tumor microenvironment is mediated through VEGF signaling pathways', Archives of Otolaryngology - Head and Neck Surgery, vol. 137, no. 11, pp. 1146-1153. https://doi.org/10.1001/archoto.2011.194
Kamat, Ameet ; Rajoria, Shilpi ; George, Andrea ; Suriano, Robert ; Shanmugam, Arul Kumaran ; Megwalu, Uchechukwu ; Prakash, Pradeep Bangalore ; Tiwari, Raj ; Schantz, Stimson. / Estrogen-mediated angiogenesis in thyroid tumor microenvironment is mediated through VEGF signaling pathways. In: Archives of Otolaryngology - Head and Neck Surgery. 2011 ; Vol. 137, No. 11. pp. 1146-1153.
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abstract = "Objectives: To explore the induction of a proangiogenic phenotype in endothelial cells in the thyroid tumor microenvironment by estrogen-treated thyroid cancer cells and to define the role of vascular endothelial growth factor (VEGF) in this interaction. Design: Cell-based in vitro systems analysis. Subjects: Thyroid tumor cell lines (BCPAP [papillary thyroid cancer] and ML-1 [follicular thyroid cancer]) were cultured with estradiol with and without an estrogen receptor (ER) inhibitor (fulvestrant or ICI) and used to treat human umbilical vein endothelial cells (HUVECs). Interventions: Immunofluorescence was used to confirm the presence of ERα and ERβ in BCPAP cells. Conditioned medium was then used to evaluate the induction of HUVEC tubulogenesis and migration. Secretion of VEGF in this medium was evaluated by Western blot analysis. The expression of phosphoinositide 3-kinase (PI3K), the initiator of a proangiogenic pathway, was evaluated with Western blot analysis of HUVEC lysates. The subsequent effects of an ER inhibitor (fulvestrant/ ICI) and a neutralizing VEGF antibody were also observed. Results: Estrogen receptor α and ERβ are expressed in thyroid cancer cells. Estrogen-stimulated ML-1 cells secreted an increased amount of VEGF likely as a result of ER signaling. In contact with this environment, HUVECs demonstrate enhanced tubulogenesis and migration. Western blot analysis documented estrogen-mediated upregulation of PI3K in HUVECs. These effects were mitigated by an ER inhibitor (fulvestrant/ICI) and a neutralizing VEGF antibody. Conclusions: Our data provide evidence that estrogen can induce a proangiogenic endothelial cell phenotype in the thyroid tumor microenvironment through ER and VEGF signaling. Our findings suggest that the effect of antiestrogenic therapy targeting tumor angiogenesis can be enhanced through VEGF inhibition.",
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AU - Kamat, Ameet

AU - Rajoria, Shilpi

AU - George, Andrea

AU - Suriano, Robert

AU - Shanmugam, Arul Kumaran

AU - Megwalu, Uchechukwu

AU - Prakash, Pradeep Bangalore

AU - Tiwari, Raj

AU - Schantz, Stimson

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N2 - Objectives: To explore the induction of a proangiogenic phenotype in endothelial cells in the thyroid tumor microenvironment by estrogen-treated thyroid cancer cells and to define the role of vascular endothelial growth factor (VEGF) in this interaction. Design: Cell-based in vitro systems analysis. Subjects: Thyroid tumor cell lines (BCPAP [papillary thyroid cancer] and ML-1 [follicular thyroid cancer]) were cultured with estradiol with and without an estrogen receptor (ER) inhibitor (fulvestrant or ICI) and used to treat human umbilical vein endothelial cells (HUVECs). Interventions: Immunofluorescence was used to confirm the presence of ERα and ERβ in BCPAP cells. Conditioned medium was then used to evaluate the induction of HUVEC tubulogenesis and migration. Secretion of VEGF in this medium was evaluated by Western blot analysis. The expression of phosphoinositide 3-kinase (PI3K), the initiator of a proangiogenic pathway, was evaluated with Western blot analysis of HUVEC lysates. The subsequent effects of an ER inhibitor (fulvestrant/ ICI) and a neutralizing VEGF antibody were also observed. Results: Estrogen receptor α and ERβ are expressed in thyroid cancer cells. Estrogen-stimulated ML-1 cells secreted an increased amount of VEGF likely as a result of ER signaling. In contact with this environment, HUVECs demonstrate enhanced tubulogenesis and migration. Western blot analysis documented estrogen-mediated upregulation of PI3K in HUVECs. These effects were mitigated by an ER inhibitor (fulvestrant/ICI) and a neutralizing VEGF antibody. Conclusions: Our data provide evidence that estrogen can induce a proangiogenic endothelial cell phenotype in the thyroid tumor microenvironment through ER and VEGF signaling. Our findings suggest that the effect of antiestrogenic therapy targeting tumor angiogenesis can be enhanced through VEGF inhibition.

AB - Objectives: To explore the induction of a proangiogenic phenotype in endothelial cells in the thyroid tumor microenvironment by estrogen-treated thyroid cancer cells and to define the role of vascular endothelial growth factor (VEGF) in this interaction. Design: Cell-based in vitro systems analysis. Subjects: Thyroid tumor cell lines (BCPAP [papillary thyroid cancer] and ML-1 [follicular thyroid cancer]) were cultured with estradiol with and without an estrogen receptor (ER) inhibitor (fulvestrant or ICI) and used to treat human umbilical vein endothelial cells (HUVECs). Interventions: Immunofluorescence was used to confirm the presence of ERα and ERβ in BCPAP cells. Conditioned medium was then used to evaluate the induction of HUVEC tubulogenesis and migration. Secretion of VEGF in this medium was evaluated by Western blot analysis. The expression of phosphoinositide 3-kinase (PI3K), the initiator of a proangiogenic pathway, was evaluated with Western blot analysis of HUVEC lysates. The subsequent effects of an ER inhibitor (fulvestrant/ ICI) and a neutralizing VEGF antibody were also observed. Results: Estrogen receptor α and ERβ are expressed in thyroid cancer cells. Estrogen-stimulated ML-1 cells secreted an increased amount of VEGF likely as a result of ER signaling. In contact with this environment, HUVECs demonstrate enhanced tubulogenesis and migration. Western blot analysis documented estrogen-mediated upregulation of PI3K in HUVECs. These effects were mitigated by an ER inhibitor (fulvestrant/ICI) and a neutralizing VEGF antibody. Conclusions: Our data provide evidence that estrogen can induce a proangiogenic endothelial cell phenotype in the thyroid tumor microenvironment through ER and VEGF signaling. Our findings suggest that the effect of antiestrogenic therapy targeting tumor angiogenesis can be enhanced through VEGF inhibition.

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