Abstract
Purpose: To evaluate the effect of synthetic polypeptide (N15P) derived from viral macrophage inflammatory protein II (vMIP-II) on the secretion of vascular endothelial growth factor (VEGF) as well as investigate the signaling pathways involved in stromal cell-derived factor-1α(SDF-1α)/CXC Chemokin Receptor 4 (CXCR4) axis-induced VEGF in glioblastoma U87 cells.Methods: Glioblastoma U87 cells were exposed to SDF-1á, N15P with various concentrations. The expression of CXCR4, SDF-1α and VEGF mRNA were assessed by RT-PCR, while expression level of VEGF was tested by ELISA and protein kinase B (Akt) phosphorylation detected by Western blot.Results: The results showed that CXCR4, SDF-1α, VEGF are expressed in human glioblastoma U87 cell lines. SDF-1α caused a dose-dependent sensitivity of cell proliferation with a maximum effect at 15 µmole/ml, while N15P decreased cell viability in U87 cells in a dose-dependent manner. SDF-1α stimulated the activation of VEGF, and N15P inhibited the activation of VEGF with or without SDF-1α stimulation. VEGF production in U87 cells was associated with Akt pathway. These changes in intracellular processes were blocked by N15P in a dose-dependent manner.Conclusion: The results suggest that N15P suppress SDF-1α/CXCR4 Mediated VEGF production through Akt signaling pathway and this may be a potent therapeutic strategy in glioblastoma.Keywords: Viral macrophage, Inflammatory protein II, Glioblastoma, CXC chemokin receptor 4, Stromal cell-derived factor-1α, Protein kinase B
Highlights
High-grade gliomas are considered to be the most common and serious primary gliomas in adults [1]
To understand the effect of N15P on tumor viability, we examined the viability of U87 cells treated with N15P by MTT assay
The results indicate that SDF1α caused a dose-dependent sensitivity of cell proliferation with a maximum effect at 15 μmol/ml in U87 cell lines (Figure 2A)
Summary
High-grade gliomas are considered to be the most common and serious primary gliomas in adults [1]. We evaluated the inhibitory effects of N15P on the production of VEGF and the role of AKT signal pathway in vitro. For RT-PCR to detect SDF-1, VEGF and CXCR4 mRNA expression, 1 μg of total RNA from U87 cells treated with different factors was subjected to RT-PCR using two-step RT-PCR kit according to the protocol described by the supplier.
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