The Study On The Mechanism Of Invasion Evoked By Glutamate In Glioblastm U251 Cells

Objective :1,To study the relationship between glutamate and the invasion of U251cells.2,To study the relationship between glutamate and the secretion of MMP-2 and MMP-9 in U251 cells.3,To study the relationship between glutamate and the expression of EMMPRIN in U251 cells.Methods:1,U251 cells were cultured in vitro, U251 cells were treated by 100μM glutamate in DMEM medium and by DMEM medium no glutamate for 24 hours, cells invasion was detected by Matrigel Transwell method.2,We cultured cells in DMEM medium containing 100μM glutamate and in DMEM medium no glutamate for 2 hours,4 hours,8 hours,12 hours and collected the supernatant, MMP-2 and MMP-9 were detected by gelatin zymography assay.3,We cultured cells in medium DMEM medium containing 100μM glutamate and in DMEM medium no glutamate for the indicated times and collected cells, the expression of EMMPRIN was detected by Western blot assay.Results:1. The number of glutamate group invasive cells(37.44±2.31)are much more than the number of control group invasive cells(25.89±3.00), The difference has statistics meaning. (P < 0.05).2. We cultured cells in DMEM medium containing 100μM glutamate and in DMEM medium no glutamate for 2 hours,4 hours,8 hours,12 hours,and we campare the quantity of MMP-2 and MMP-9 secreted by glutamate group cells with the quantity of MMP-2 and MMP-9 secreted by control group cells,we find that: the quantity of MMP-2 secreted by glutamate group cells and control group cells have no difference from 2 to 4 hours, the quantity of MMP-2 secreted by glutamate group cells are much more than the quantity of MMP-2 secreted by control group cells from 8 to 12 hours(P<0.05); the quantity of MMP-9 secreted by glutamate group cells and control group cells have no difference from 2 to 12 hours.3. We cultured cells in DMEM medium containing 100μM glutamate and in DMEM medium no glutamate for 3 hours,6 hours,9 hours ,the expression of EMMPRIN have no Significant difference between the glutamate group cells and the control group cells from 3 to 9 hours.Conclusion:Glutamate promote U251 cells to invade ; glutamate promote U251 cells to secrete MMP-2, MMP-2 can degrade extracellular matrix thereby enhance the invasion of glioma cells, MMP-9 and MMP-2 have similar function,but glutamate do not promote U251 cells to secrete MMP-9,so that glutamate promote U251 cells to secrete MMP-2 is one of aspects which glutamate promote U251 cells to invade; EMMPRIN can promote glioma cells to secrete MMP-2 ,were not involved in that glutamate promoted invading of U251 cells,that glutamate promote U251 cells to secrete MMP-2 hasn't relationship with EMMPRIN .