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I Notting, B Sijmons, HM H Hurks, G van der Pluijm, MJ Jager; Expression Of The Vegf Family And Its Receptors Of 12 Uveal Melanoma Cell Lines With Pcr In Vitro . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1116.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose:Angiogenesis is an important process in the dissemination of uveal melanoma stimulated by different angiogenic factors. We investigated the presence of vascular endothelial growth factor (VEGF) A, B, C, D, and their receptors Flt1, Flt4 and KDR with RT-PCR in 12 uveal melanoma cell lines. Methods:We used eight cell lines (92-1, Mel-202, -270, -285, -290, OCM-1, -3, -8) of primary uveal melanomas and one cell line, one, OMM-1, of a uveal melanoma skin metastasis. Cell line Mel 270 was derived from the primary tumor of a patient. Two cell lines, OMM 1.3 and 1.5, came from liver metastasis of the same patient. RNA has been isolated from cells in culture according to the method described by Chomcynski and Sacchi. CDNA was co amplified with human specific beta 2-microglobulin primers over 38 cycles in the presence of 4-fold serial dilutions of the internal standards pQA1. Negative controls were run in parallel in each experiment. Aliquots of 15ml of each amplified sample and 100 base pair DNA ladder were subjected to electrophoresis on agarose gel and were photographed. We analyzed the intensity of the PCR results with Scion Image. Human-specific oligonucleotide primers for VEGF A, B, C, D and their receptors KDR, FLT1, FLT4 were used. Results:We observed high expression of VEGF B, C and their receptors KDR and FLT4 in all 12 uveal melanoma cell lines. KDR and Flt4 expression varied between the cell lines. VEGF A was expressed only in three primairy uveal melanoma cell lines and two metastatic cell lines. There was no expression of VEGF D and Flt1 in any cell line. Conclusion:Uveal melanoma cell lines expressed high levels of VEGF B, C, KDR and FLT4 in vitro. VEGF-A may not be an important factor in the angiogenesis of uveal melanoma. The role of VEGF B and C could be of great importance in the angiogenesis of uveal melanoma.
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