The Effect Of Endostatin On Glioma Cell Proliferation

Glioma derived from the neural epithelium is the most common primary intracranial tumor, with a high incidence of recurrence rate, high mortality rate and low cure rate. With the current technical conditions, the prognosis of glioma, especially highly malignant glioma, is far from ideal. At present, more and more people pay attention to the anti-angiogenic therapy, and the anti-angiogenic therapy that inhibit proliferation of tumor cells by inhibiting angiogenesis and access to nutrition, to achieve the inhibition effect of tumor growth and metastases. Recombinant human endostatin (abbreviation endostatin) is an antineoplastic in clinical with mainly used for the initial treatment or retreatment Ⅲ/Ⅳ of non-small cell lung cancer, But how the treatment for glioma is not clear. Therefore, this experiment intends to evaluate the effect of recombinant human endostatin on glioma lines and the expression of VEGF and Nucleolin, and further to provide a new approach and theoretical basis for the clinical treatment of brain glioma.PART ONE The Endostatin Inhibits the Proliferation of Glioma CellsObjective To study the inhibition of proliferation and migration of endostatin on glioma cell lines (U251and C6), and to investigate treatment and significance of endostatin treatment gliomas.Methods U251and C6cells were cultured in vitro, and different concentrations of endostatin(10,20,30,40,50μg/ml) treat with U251and C6for48h. Then MTT assay the inhibition effect of cells proliferation of different concentrations of endostatin; and20μg/ml endostatin treat with cells for24and48hours, to observe the affect of cells migration and the morphological changes.Results Different concentrations of endostatin (10,20,30,40,50μg/ml) on the proliferation of U251and C6cells have different degrees of inhibition, and select the20μg/ml concentration of endostatin as the best concentration. In addition, the migration of U251and C6cells after scratches are significantly inhibited by endostatin. Conclusion Endostatin can inhibit the proliferation and migration of glioma cells in vitro. PART TWO The Effect of Endostatin on the Expression of VEGF and Nucleolin of Glioma CellsObjective Investigate the expression of VEGF and nucleolin protein of U251and C6cells before and after the endostatin treatment. Further to provide the experimental basis and the scientific and theoretical basis for the clinical treatment of endostatin on tumor.Methods Using immunofluorescence mark U251and C6cells, and observe the expression of VEGF and Nucleolin protein and the changes of expression of VEGF after20μg/ml endostatin treatment for24and48h, then using western blot analysis the expression changes of nucleolin protein of two kinds of cells after20μg/ml endostatin treatment for different hours.Results VEGF are highly expressed in two kinds of cells, mainly distributed in the cytoplasm; Nucleolin was also expressed in two kinds of cells, the main expression in the nucleus, a small amount of expression in the cytoplasm and cell membrane. Endostatin can down-regulation expression of VEGF of U251and C6cells; while the expression of nucleolin is decreased first and then increased.Conclusion Endostatin can down-regulation the expression of VEGF in glioma cells in vitro; and can affect the expression of nucleolin. Speculating that VEGF and nucleolin play a role in endostatin inhibition proliferation of glioma cells, but its specific mechanism of action is not yet clear.