Contributions
Abstract: PB2091
Type: Publication Only
Background
Myelodysplastic syndromes (MDS) is a heterogeneous group of hematological disorders characterized by impaired hematopoiesis with a high risk of leukemic transformation. Recently, overexpression of Vascular endothelial growth factor (VEGF), a major angiogenic factor, has been found in MDS and showed different expression status in different risk of MDS.
Aims
Our aim is to investigate the possible role of miR-15a and miR-16 on regulation of VEGF expression and their effects on angiogenesis in different risk of MDS.
Methods
ELISA, immunohistochemistry assay and immunofluorescence were used for detection of VEGF secretion in peripheral blood of MDS patients, protein expression in biopsies of MDS patients and several leukemia and MDS cell lines, repectively. qRT-PCR was performed to detect the expression of miR-15a and miR-16 in MDS bone marrow samples and cell lines. A human MDS SKM-1 cell line was transfected with pre-miR-16, anti-miR-16 and their respective negative control. ELISA and western blotting were performed to test the effect of miR-16 on the secretion and protein expression of VEGF. The luciferase reporter assays was conducted to confirm VEGF is a potential target of miR-16. The migration and tube formation of human umbilical vein endothelial cells were performed in miR-16 transfected SKM-1 condition medium (CM) .
Results
Levels of miRNA-16 were significantly decreased in high-risk MDS patients as well as SKM-1 cell line, while the expression levels of VEGF were upregulated. Transfection of miR-16 in SKM-1 cells resulted in reduced secretion and protein expression of VEGF. The migration and tube formation of human umbilical vein endothelial cells were decreased in miR-16 transfected SKM-1 CM compared to CM from SKM-1 cells transfected with vector control. Direct binding of miR-16 to the 3’ untranslated region of VEGF was confirmed by luciferase reporter assay.
Conclusion
These data suggest that miR-16 may play a role in disease progression of MDS partially by modulation of angiongenesis through targeting VEGF and that miR-16 might serve as a novel therapeutic target in MDS.
Session topic: 10. Myelodysplastic syndromes – Clinical
Keyword(s): Angiogenesis, Myelodysplasia, VEGF
Abstract: PB2091
Type: Publication Only
Background
Myelodysplastic syndromes (MDS) is a heterogeneous group of hematological disorders characterized by impaired hematopoiesis with a high risk of leukemic transformation. Recently, overexpression of Vascular endothelial growth factor (VEGF), a major angiogenic factor, has been found in MDS and showed different expression status in different risk of MDS.
Aims
Our aim is to investigate the possible role of miR-15a and miR-16 on regulation of VEGF expression and their effects on angiogenesis in different risk of MDS.
Methods
ELISA, immunohistochemistry assay and immunofluorescence were used for detection of VEGF secretion in peripheral blood of MDS patients, protein expression in biopsies of MDS patients and several leukemia and MDS cell lines, repectively. qRT-PCR was performed to detect the expression of miR-15a and miR-16 in MDS bone marrow samples and cell lines. A human MDS SKM-1 cell line was transfected with pre-miR-16, anti-miR-16 and their respective negative control. ELISA and western blotting were performed to test the effect of miR-16 on the secretion and protein expression of VEGF. The luciferase reporter assays was conducted to confirm VEGF is a potential target of miR-16. The migration and tube formation of human umbilical vein endothelial cells were performed in miR-16 transfected SKM-1 condition medium (CM) .
Results
Levels of miRNA-16 were significantly decreased in high-risk MDS patients as well as SKM-1 cell line, while the expression levels of VEGF were upregulated. Transfection of miR-16 in SKM-1 cells resulted in reduced secretion and protein expression of VEGF. The migration and tube formation of human umbilical vein endothelial cells were decreased in miR-16 transfected SKM-1 CM compared to CM from SKM-1 cells transfected with vector control. Direct binding of miR-16 to the 3’ untranslated region of VEGF was confirmed by luciferase reporter assay.
Conclusion
These data suggest that miR-16 may play a role in disease progression of MDS partially by modulation of angiongenesis through targeting VEGF and that miR-16 might serve as a novel therapeutic target in MDS.
Session topic: 10. Myelodysplastic syndromes – Clinical
Keyword(s): Angiogenesis, Myelodysplasia, VEGF