| In the central nervous system glioma is the most common and most difficult to cure cancer, Neuroectodermal origin of malignant primary CNS tumors, accounting for 35.26%-60.96% of intracranial tumors,and accounting for all adults 2% of malignant tumors,is a common disease to human health that has serious harm. In the development of,it having invasive growth,aggressive,easy to relapse,rich in blood vessels and other biological characteristics,and is generally believed that,the characteristic of glioma and its relationship to vascular endothelial cell cause gliomas difficult to cure,the prognosis is poor,easy to relapse. The interaction between glioma cells and vascular endothelial cells may promote the invasive growth of glioma, but the specific mechanism is not very clear. Therefore, the study of human brain microvascular endothelial cells and glioma cells interactions between cells have a very important practical significance. As the most common malignancy in humans, glioma is essentially a multi-gene participation, and multi-gene abnormality disease. Related findings show, ERK / MAPK signal transduction pathway is one of the main signal transduction pathway in glioma, and is currently associated with the pathogenesis, development and biological behavior of glioma. In the present study, in order to explore the related mechanisms between glioma cells and endothelial cells, we observed EGF secreted by the vascular endothelial cells on biological behavior of glioma cell proliferation and invasion capacity, observed the related factors in turn affect the ERK signaling pathway.Contents and MethodsVascular endothelial cells secrete Factor EGF may affect neurobiology behavior of glioma cells. Cultured glioma cell line U87MG?U251, and microvascular endothelial cells(HBMEC), establish co-cultured glioma cells and vascular endothelial cells between the two systems; join ERK signaling pathway agonist EGF and inhibitor PD98059 treatment of human EGF when glioma,observing glioma cell proliferation, invasion and Vascular morphology phenomenon when ERK activity changed. The cells are grouped according to the experimental requirements, glioma cells were cultured alone, the two co-culture group, added EGF group and PD98059 group. They were taken CCK-8 assay the ability of value-added glioma cells, Transwell invasion assay invasion of glioma cells, synthetic vascular morphology assay glioma cells Vascular morphogenesis, Western Blots law detect glioma cells P-ERK activation.Investigate the effects of VEGF to biological behavior and EGF secretion of endothelial cells that secreted by glioma cells and. The endothelial cells are grouped according to the experimental requirements, HBMEC cultured alone, HBMEC and glioma cells co-cultured group. ELISA assay the secretion of EGF at different conditions in vascular endothelial cells, angiogenesis assay the HBMEC angiogenesis under different case.Result:(1) The results showed that, compared with the control group, co-cultured with HBMEC glioma cells proliferation ability significantly enhanced. When added to 50 ng / m L exogenous EGF, proliferation was significantly enhanced. And joining 5?M / L PD98059, value-added significantly reduced.(p <0.05)(2) Compared with the control group(number invasion was 105.33 ± 6.110), co-culture group U87 MG invasive ability was significantly enhanced(the number of invasion was 261.67 ± 10.214), U87 MG added with 50 ng / m L EGF passage cells was significantly enhanced( the number of invasion was 214.33 ± 11.150). The U87 MG added 5?M / L PD98059 cell invasion was significantly reduced(the number of invasion was 55.67 ± 6.028). U251 cells also showed similar results, the control group, co-culture group, EGF group added, adding the number of cell invasion PD98059 group were: 77.33 ± 6.658, 237.00 ± 10.440,191.33 ± 3.215,50.67 ± 4.509.( p <0.05)(3) Compared with control group(luminal diameter: 130.112 ± 2.022?m), co-culture group U87 MG merging connected into a network-like structure, the ability of form blood vessel formation was significantly enhanced(202.134 lumen diameter ± 2.007?m), U87 MG join 50 ng / m L EGF group(223.19 ± 2.012?m) vascular morphogenesis quasi ability significantly enhanced, and added 5?M / L PD98059 group(76.199 ± 2.833?m) cell growth disorder, to be significantly smaller lumen vascular morphology. U251 cells also showed similar results, the control group, co-culture group, add EGF group, add PD98059 group the number cell invasion were: 90.122 ± 2.733?m 230.112 ± 2.662?m, 143.2262.098?m 44.039 ± 1.092?m.(p <0.05)(4) GAPDH as internal control, the results show, P-ERK expression of co-culture group was significantly enhanced. Join 50 ng / m LEGF group P-ERK expression is relatively enhanced. Join 5u M / L PD98059 group P-ERK expression was significantly reduced. p <0.05(5) The results showed that when join different concentrations of VEGF165 to HBMEC, the secretion of EGF continue increase. When co-cultured with the two glioma cells, EGF secretion increased significantly.(p <0.05)(6) The results showed that compared with the control group(luminal diameter: 115.12 ± 1.731?m),the ability of the angiogenesis was significantly increased when co-cultured, mutual integration between cells, the formation of the network structure, and the number of small tubes and pipes luminal diameter increased significantly(luminal diameter: 245.228 ± 1.993?m). U251 cells also showed similar results(luminal diameter: 225.163 ± 1.622?m).( p <0.05)Conclusion:1. The growth factors EGF that secreted by vascular endothelial cells might increase the proliferation ability ?invasive ability and synthetic vascular morphogenesis of glioma cells by RAF-MAPK-ERK signaling pathway, P-ERK expression was also increased.2. The VEGF secreted by Glioma cells might increase the EGF secretion of vascular endothelial cells by RAF-MAPK-ERK signaling pathway,and increasing the ability of angiogenesis. |
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