Regulation of VEGF-A, VEGFR-I, Thrombospondin-1, -2, and -3 Expression in a Human Pituitary Cell Line (HP75) by TGFβ1, bFGF, and EGF

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Pituitary tumors are highly vascular neoplasms, which suggest an important rol of angiogenesis in pituitary tumor growth. We used the human pituitary cell line (HP75) to examine the effects of the growth factors TGFβ1, bFGF, and EGF on cell growth, and on the regulation of the pro-angiogenic growth factor VEGFA and the VEGFR-I and the anti-angiogenic molecules thrombospondin (TSP) TSP-1 and TSP-2 along with TSP-3. Real-time RT-PCR was used to measure mRNA levels, and Western blot was used to analyze TSP-1 and TSP-2 protein levels.TGFβ1 treatment (1 × 10−9M) increased VEGF-A mRNA levels significantly (p < 0.05) after 4 and 24 h of treatment. TGF β1 treatment decreased VEGF-R mRNA levels after 96 h of treatment (p < 0.05). After 96 h of treatment, TSP-1 and TSP-2 mRNA levels were significantly increased (p < 0.05) by TGFβ1 treatment, which also inhibited HP75 cell growth. Basic FGF also increased TSP-1 mRNA levels after 96 h of treatment, but did not regulate growth of the pituitary tumor cells. Basic FGF and EGF did not modulate changes in VEGF-A mRNA levels after 4 and 24 h of treatment, but EGF increased VEGF-A significantly (p < 0.05) after 96 h of treatment.These results indicate that TGFβ1 treatment may regulate angiogenesis in pituitary cells by initially increasing levels of pro-angiogenic VEGF-A and then stimulating the anti-angiogenic molecules TSP-1 and TSP-2 levels.

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