ABSTRACT
Neuroblastoma(NB)is the most common extracranial solid tumor derived from the sympathetic ner- vous system in childhood. The location of NB is variable, the pathogenesis is complex and the clinical manifestations vary greatly. The prognosis is good in low or medium risk NB patients but bad in high risk NB patients. Recent studies have shown that the occurrence, growth, proliferation, migration and invasion of NB as well as the prognosis of NB pa- tients are associated with the abnormal expression of signaling proteins in NB, including the transcriptional regulators, kinases, and receptors, etc.. This article reviews the role and action mechanism of different signaling proteins in the oc- currence and development of NB.
ABSTRACT
PURPOSE: alpha-Galactosylceramide (alpha-GalCer)-stimulated human Valpha24 natural killer T (NKT) cells exert antitumor activity against some leukemia in a CD1d dependent and TCR-mediated manner, but could not kill CD1d-negative neuroblastoma (NB) cells. There are few reports about the direct antitumor effect of highly secreted cytokines by these cells on activation. In this study, using a cell-free supernatant (SPN) collected from plate bound hCD1d/alphaGalCer tetramers-stimulated NKT cells, we examined whether they could be helpful in the immunotherapeutic treatment of NB. METHODS: Cells were cultured in IMDM. The cytokines produced by NKT cells were measured with Cytometric Bead Array (CBA) analysis. Cell viability was evaluated by calcein-AM fluorescence with digital image microscopy scanning (DIMSCAN). The percentage of specific apoptosis was calculated by flow cytometric detection of apoptosis using annexin V and 7-AAD. RESULTS: The activated NKT cells secreted high levels of IL-2, INF-gamma, TNF-alpha. The SPN was significantly cytotoxic against four out of eight tested NB cell lines, through mainly apoptosis as evidenced by annexin-V staining and inhibition with the pretreatment of pancaspase blocker. This apoptosis was significantly inhibited when anti-TNF-alpha and anti-IFN-gamma neutralizing mAbs were used separately and it was completely abolished when the two mAbs were combined. CONCLUSION: IFN-gamma and TNF-alpha produced by NKT cells could exert synergistically direct anti-tumor activity through apoptosis on some NB cell lines.
Subject(s)
Humans , Annexin A5 , Apoptosis , Cell Line , Cell Survival , Cytokines , Fluorescence , Interleukin-2 , Leukemia , Microscopy , Natural Killer T-Cells , Neuroblastoma , Tumor Necrosis Factor-alphaABSTRACT
Objective The present study was designed to explore the inhibitory effect of antisense VEGF-(165) cDNA on angiogenesis,growth rate of human neuroblastoma.Methods The eukaryotic expression vectors bearing antisense VEGF-(165) cDNA or sense VEGF-(165) cDNA were constructed.The stable cell lines transfected with the sense or antisense VEGF-(165) cDNA were established by using the selective medium containing 400?mg/L of G418.These cell lines were further studied for the exogenous antisense VEGF mRNA expression by RT-PCR and inhibition of expression of endogenous VEGF protein by immunocytochemical staining and ELISA.The proliferation of the transfected cells was analyzed by MTT method.Transfected cells were subcutaneously transplanted into nude mice and the growth of tumor masses was observed and weighted.Results The antisense VEGF was only detected in SH-SY5Y/AsVEGF cells by RT-PCR.The expression of VEGF protein was dramatically declined in the SH-SY5Y/AsVEGF cells compared with the original cells and empty vector transfected cells by immunocytochemical staining and ELISA.There was no effect of antisense VEGF transfection on cell proliferation in vitro.The growth of tumor mass with the cells transfected with antisense VEGF was significantly decreased in nude mice.Conclusion Antisense VEGF cDNA transfection to SH-SY5Y cells can effectively inhibit the expression of endogenous VEGF protein and tumor growth in nude mice.