The Study Of The Effects Of The Rhe Roles Of LDHA On The Growth And EMT In Glioma Cells

Objective : To investigate the roles of LDHA in the growth and epithelial-mesenchymal transition in glioma cells,which might be used to develop validated sets of clinical diagnostic and diagnostic for glioma.Methods:(1)qRT-PCR and western blot were used to determine LDHA mRNA and protein levels in three glioma cells,respectively;(2)U87MG?U251MG and SW1783 cells were transfected with sh-LDHA plasmids,and then verifiying the Transfection efficiency;(3)The CCK8 and colony formation assaies were applied to assess the proliferative capacities of glioma cells;(4)The flow cytometry were performed to examine the apoptosis of U87 MG cells;(4)The wound healing assay and transwell migration assay were conducted to evaluate the migration abilities of glioma cells;(5)Lactate dehydrogenase activity was determined by an assay kit The lacate dehydrogenase assay kit;(6)The western blot were used to analyze the EMT related proteins expression after knocking down LDHA in glioma cells.Results:(1)The expression prolifes of LDHA were greatly varied in glioma cells,and the expression levels of LDHA in U87 MG cells were higher than the other two glioma cells;(2)LDHA Konckdown has no effects on the proliferation of U251 MG cells,and the decreased proliferation in U251 MG cells after knocking down LDHA was not significant.While,it showed that LDHA knockdown dramatically inhibited the proliferation of U87 MG cells;(3)LDHA knockdown decrased the migration of U87 MG cells;(4)LDHA knockdown induced the apoptosis of U87 MG cells;(5)LDHA knockdown reduced the lactate dehydrogenase activity in U87 MG cells;(6)The results turned that the epithelial marker E-cadherin expression was enhanced,while the expression of mesenchymal markers Fibronectin,N-cadherin,Vimentin and snail were lower after inhibiting LDHA expression,the EMT was inhibited.However,there were few effects on the EMT in SW1783 cells and no effects in U251 MG cells.Conclusions:(1)LDHA knockdown inhibited the proliferation and migration,and induced the apoptosis in U87 MG cells;(2)LDHA knockdown could reverse the EMT in U87 MG cells.