Interaction between VEGF and calcium-independent phospholipase A2 in proliferation and migration of retinal pigment epithelium.
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Interaction between VEGF and calcium-independent phospholipase A2 in proliferation and migration of retinal pigment epithelium. / Kehler, AK; Andersen, C; Andreasen, JR; Vohra, Rupali; Junker, N; Poulsen, KA; Kolko, Miriam.
In: Current Eye Research, 01.06.2012.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Interaction between VEGF and calcium-independent phospholipase A2 in proliferation and migration of retinal pigment epithelium.
AU - Kehler, AK
AU - Andersen, C
AU - Andreasen, JR
AU - Vohra, Rupali
AU - Junker, N
AU - Poulsen, KA
AU - Kolko, Miriam
PY - 2012/6/1
Y1 - 2012/6/1
N2 - PurposeInhibition of VEGF in the eye is an important treatment modality for reducing proliferation and migration of retinal pigment epithelium (RPE) in age-related macular degeneration (AMD). Additionally, previous studies suggest calcium-independent phospholipase A(2) group VIA (iPLA(2)-VIA) to be a potential regulator of cell proliferation and migration, and evidence show abundant expression of iPLA(2)-VIA in RPE cells. The aim of the present study was to evaluate the potential role of iPLA(2)-VIA in VEGF-induced proliferation and migration of RPE cells.Materials and methodsThe human RPE cell line, ARPE-19, was used in all assays. To explore the role of iPLA(2)-VIA in VEGF-induced RPE proliferation and migration, iPLA(2)-VIA inhibition by the iPLA(2)-VIA specific inhibitor, bromoenol lactone, was done. RPE cell proliferation and migration were evaluated by measurements of incorporated radioactive thymidine in DNA and by a Boyden chamber technique, respectively. A luciferase assay monitored the VEGF-induced iPLA(2)-VIA transcriptional activity. Western blot analysis and an activity assay were used to detect the protein levels and activity of iPLA(2)-VIA respectively after treatment with VEGF.ResultsRPE cells treated with VEGF showed significant increased proliferation and migration. Furthermore, inhibition of iPLA(2)-VIA significantly reduced the spontaneous proliferation and migration as well as the VEGF-induced proliferation and migration. Finally, inhibition of iPLA(2)-VIA reduced the VEGF-induced iPLA(2)-VIA-activity, -protein level, and -promoter activity.ConclusionsA significant interaction between VEGF and iPLA(2)-VIA in the regulation of RPE cells appears to be relevant in elucidating the exact mechanisms of action in the proliferative and migratory phenotype of RPE cells in AMD.
AB - PurposeInhibition of VEGF in the eye is an important treatment modality for reducing proliferation and migration of retinal pigment epithelium (RPE) in age-related macular degeneration (AMD). Additionally, previous studies suggest calcium-independent phospholipase A(2) group VIA (iPLA(2)-VIA) to be a potential regulator of cell proliferation and migration, and evidence show abundant expression of iPLA(2)-VIA in RPE cells. The aim of the present study was to evaluate the potential role of iPLA(2)-VIA in VEGF-induced proliferation and migration of RPE cells.Materials and methodsThe human RPE cell line, ARPE-19, was used in all assays. To explore the role of iPLA(2)-VIA in VEGF-induced RPE proliferation and migration, iPLA(2)-VIA inhibition by the iPLA(2)-VIA specific inhibitor, bromoenol lactone, was done. RPE cell proliferation and migration were evaluated by measurements of incorporated radioactive thymidine in DNA and by a Boyden chamber technique, respectively. A luciferase assay monitored the VEGF-induced iPLA(2)-VIA transcriptional activity. Western blot analysis and an activity assay were used to detect the protein levels and activity of iPLA(2)-VIA respectively after treatment with VEGF.ResultsRPE cells treated with VEGF showed significant increased proliferation and migration. Furthermore, inhibition of iPLA(2)-VIA significantly reduced the spontaneous proliferation and migration as well as the VEGF-induced proliferation and migration. Finally, inhibition of iPLA(2)-VIA reduced the VEGF-induced iPLA(2)-VIA-activity, -protein level, and -promoter activity.ConclusionsA significant interaction between VEGF and iPLA(2)-VIA in the regulation of RPE cells appears to be relevant in elucidating the exact mechanisms of action in the proliferative and migratory phenotype of RPE cells in AMD.
UR - https://doi.org/10.3109/02713683.2012.663855
U2 - 10.3109/02713683.2012.663855
DO - 10.3109/02713683.2012.663855
M3 - Tidsskriftartikel
C2 - 22577768
JO - Current Eye Research
JF - Current Eye Research
SN - 0271-3683
ER -
ID: 336754940