Journal:

Article Title: 2-Methoxyestradiol Exhibits a Biphasic Effect on VEGF-A in Tumor Cells and Upregulation Is Mediated Through ER-?: A Possible Signaling Pathway Associated with the Impact of 2-ME 2 on Proliferative Cells 1

doi:

Figure Lengend Snippet: Dose-dependent effects of 2-ME2 on cell proliferation in rat pituitary tumor-derived GH3 tumor cells, human breast tumor-derived MCF-7 tumor cells, and human pancreatic adenocarcinoma MIA-PaCa-2 cells. Exponentially growing cells in DMEM containing 10% serum were exposed to various concentrations of 2-ME2 for 3 days, and cell proliferation was determined by measuring the [3H]thymidine incorporation into DNA. (A) GH3 rat pituitary tumor cells. (B) MCF-7 human breast tumor cells. (C) MIA-PaCa-2 pancreatic adenocarcinoma cells. Data displayed as mean ± SD from three separate experiments. P value was determined by Student's t-test. *P < .05 vs control; **P < .01 vs control; ***P < .001 vs control.

Article Snippet: Cell Lines and Culture Conditions GH3 rat pituitary tumor cells, human breast tumor-derived MCF-7 epithelial tumor cells, and MIA-PaCa-2 pancreatic adenocarcinoma cells were obtained from the American Type Cell Culture Collection (ATCC; Rockville, MD).

Techniques: Derivative Assay, Control

Journal:

Article Title: 2-Methoxyestradiol Exhibits a Biphasic Effect on VEGF-A in Tumor Cells and Upregulation Is Mediated Through ER-?: A Possible Signaling Pathway Associated with the Impact of 2-ME 2 on Proliferative Cells 1

doi:

Figure Lengend Snippet: Dose-dependent effects of 2-ME2 on VEGF-A mRNA expressions in GH3 rat pituitary tumor cells, MCF-7 human breast tumor cells, and MIA-PaCa-2 human pancreatic adenocarcinoma cells. Exponentially growing cells in DMEM containing 10% serum were exposed to various concentrations of 2-ME2 for 3 days, and VEGF-A mRNA expression was determined by Northern blot analyses using nonradioactive DIG-labeled probe. (A), (C), and (E) represent VEGF-A and GAPDH mRNA levels in untreated and treated GH3, MCF-7, and MIA-PaCa-2 cells respectively. (B), (D), and (F) represent arbitrary values indicating VEGF-A mRNA levels. Data displayed as mean±SD from three separate experiments. P value was determined by Student's t-test. *P < .05 vs control; **P < .01 vs control.

Article Snippet: Cell Lines and Culture Conditions GH3 rat pituitary tumor cells, human breast tumor-derived MCF-7 epithelial tumor cells, and MIA-PaCa-2 pancreatic adenocarcinoma cells were obtained from the American Type Cell Culture Collection (ATCC; Rockville, MD).

Techniques: Expressing, Northern Blot, Labeling, Control

Journal:

Article Title: 2-Methoxyestradiol Exhibits a Biphasic Effect on VEGF-A in Tumor Cells and Upregulation Is Mediated Through ER-?: A Possible Signaling Pathway Associated with the Impact of 2-ME 2 on Proliferative Cells 1

doi:

Figure Lengend Snippet: Chronic effects of various doses of 2-ME2 on the levels of VEGF-A protein in different tumor cell lines. Exponentially growing cells (i.e., GH3 or MCF-7 or MIA-PaCa-2 cells) in DMEM containing 10% serum were exposed to one of the three concentrations (i.e., 1.0, 5.0, and 10.0 µM) of 2-ME2 for 3 days. After exposure, proteins were extracted from different cells and 50 µg of total cell lysates was prepared for Western immunoblot analysis. VEGF-A protein levels were determined by chemiluminescent immunoblotting with monoclonal anti-human VEGF-A. The photograph exhibits a single representative blot showing VEGF-A and β-actin levels in untreated and 2-ME2-treated cells.

Article Snippet: Cell Lines and Culture Conditions GH3 rat pituitary tumor cells, human breast tumor-derived MCF-7 epithelial tumor cells, and MIA-PaCa-2 pancreatic adenocarcinoma cells were obtained from the American Type Cell Culture Collection (ATCC; Rockville, MD).

Techniques: Western Blot

Journal:

Article Title: 2-Methoxyestradiol Exhibits a Biphasic Effect on VEGF-A in Tumor Cells and Upregulation Is Mediated Through ER-?: A Possible Signaling Pathway Associated with the Impact of 2-ME 2 on Proliferative Cells 1

doi:

Figure Lengend Snippet: Time- and dose-dependent effects of 2-ME2 on VEGF-A mRNA expression in GH3 cells. Semiconfluent GH3 cells were exposed to various doses of 2-ME2 (i.e., 1, 5, and 10 µM) for different times as indicated; 0.1% ethanol-treated culture was considered as the untreated control (C). After treatment, total RNA was extracted and VEGF-A mRNA expression was determined by Northern blot analyses using nonradioactive DIG-labeled probe. Data displayed as mean ± SD from three separate experiments. (A) Single representative blot showing VEGF-A and GAPDH expressions in untreated and 2-ME2-treated cells. (B) Arbitrary values indicate VEGF-A mRNA level. Data displayed as mean±SD in each case. P value was determined by Student's t-test. P < .01 compared to untreated control.

Article Snippet: Cell Lines and Culture Conditions GH3 rat pituitary tumor cells, human breast tumor-derived MCF-7 epithelial tumor cells, and MIA-PaCa-2 pancreatic adenocarcinoma cells were obtained from the American Type Cell Culture Collection (ATCC; Rockville, MD).

Techniques: Expressing, Control, Northern Blot, Labeling

Journal:

Article Title: 2-Methoxyestradiol Exhibits a Biphasic Effect on VEGF-A in Tumor Cells and Upregulation Is Mediated Through ER-?: A Possible Signaling Pathway Associated with the Impact of 2-ME 2 on Proliferative Cells 1

doi:

Figure Lengend Snippet: Effect of pure antiestrogen ICI 182,780 on 2-ME2-induced upregulation of VEGF-A mRNA expression in GH3 cells. (A) GH3 cells were exposed to 1 µM 2-ME2 alone or in combination with 1 µM ICI 182,780 for 3 days. Total RNA was isolated and analyzed by Northern blotting using nonradioactive DIG-labeled PCRgenerated probes for VEGF-A and GAPDH. (B) The arbitrary values indicate VEGF-A mRNA concentrations. Data displayed as mean ± SD from three separate experiments. P value was determined by Student's t-test. *P < .01 vs control.

Article Snippet: Cell Lines and Culture Conditions GH3 rat pituitary tumor cells, human breast tumor-derived MCF-7 epithelial tumor cells, and MIA-PaCa-2 pancreatic adenocarcinoma cells were obtained from the American Type Cell Culture Collection (ATCC; Rockville, MD).

Techniques: Expressing, Isolation, Northern Blot, Labeling, Control

Journal:

Article Title: 2-Methoxyestradiol Exhibits a Biphasic Effect on VEGF-A in Tumor Cells and Upregulation Is Mediated Through ER-?: A Possible Signaling Pathway Associated with the Impact of 2-ME 2 on Proliferative Cells 1

doi:

Figure Lengend Snippet: Effects of anti-VEGF antibody and recombinant VEGF protein on the action of 2-ME2 on the proliferation of GH3 and MCF-7 cells. Exponentially growing cells in DMEM containing 10% serum were exposed to various concentrations of 2-ME2 in the presence or absence of VEGF antibodies or recombinant VEGF protein for 3 days, and then cell proliferation was determined using BrdU ELISA assay. Data displayed as mean ± SD from three separate experiments. P value was determined by Student's t-test. *P < .05 vs control; **P < .01 vs 2-ME2-treated cells.

Article Snippet: Cell Lines and Culture Conditions GH3 rat pituitary tumor cells, human breast tumor-derived MCF-7 epithelial tumor cells, and MIA-PaCa-2 pancreatic adenocarcinoma cells were obtained from the American Type Cell Culture Collection (ATCC; Rockville, MD).

Techniques: Recombinant, Enzyme-linked Immunosorbent Assay, Control

Journal:

Article Title: 2-Methoxyestradiol Exhibits a Biphasic Effect on VEGF-A in Tumor Cells and Upregulation Is Mediated Through ER-?: A Possible Signaling Pathway Associated with the Impact of 2-ME 2 on Proliferative Cells 1

doi:

Figure Lengend Snippet: Effect of 2-ME2-induced VEGF-A in GH3 cells on neighboring endothelial cell proliferation. The effect was determined by indirect coculture using Transwell culture system. The detailed experimental procedure has been described in the Materials and Methods section. Endothelial cell proliferation was determined by measuring the radioactive thymidine incorporation using a liquid scintillation counter. Data displayed as mean±SD from three separate experiments. P value was determined by Student's t-test. *P < .05 vs untreated control; **P < .01 vs 2-ME2-treated cells.

Article Snippet: Cell Lines and Culture Conditions GH3 rat pituitary tumor cells, human breast tumor-derived MCF-7 epithelial tumor cells, and MIA-PaCa-2 pancreatic adenocarcinoma cells were obtained from the American Type Cell Culture Collection (ATCC; Rockville, MD).

Techniques: Control