Effect Of HMGA1 On Glycolytic Pathways Of T47D Cells

Objective: To research the impact of HMGA1 in the glycolysis metabolic pathways in T47 D cells and to discuss the pathogenic mechanism.Methods: T47 D cell line were cultured in vitro.After transient transfection by using Lipofectamine,the group was divided into three groups:HMGA1,pc DNA3.1 and T47 D.Five parts as follows: 1.T47 D cells were transfected HMGA1 plasmid by using lipofectamine.2.Exploring the effect of HMGA1 on the enzyme such as hexokinase,pyruvate dehydrogenase and so on in T47 D cell by Western Blotting.3.Detecting the glucose consumption,accumulation of lactic acid and the generation of ATP in three different group by enzyme assay to further discuss the effect of HMGA1 on glycolytic pathway.4.Through detecting the activity of glycolytic key enzyme HK?PK ?PDHK?LDH and PDH to furtherly defined HMGA1 effects on glycolytic pathway.5.The stable cells of overexpresson HMGA1 and empty plasmid cells were determined by MTT assay and plate clone formation assay to test the influence between HMGA1 and cell reproductive capacity.Results: 1.Western Blot showed that,the protein of Hexokinase?Pyruvate Kinase2 ? PDH Kinase ? Phosphofrutokinase ? lactic dehydrogenase were highly expressed.But the protein expression of PDH was reduced.There were no obviously changes between HK1?PKM1/2 protien.2.The glucose consumption,accumulation of lactic acid and production of ATP was increased in HMGA1 group.3.The result showed that the enzymatic activity of PKM?KH?LDH?PFK?PDHK were significantly enhanced in HMGA1 group.4.By comparing the cells between pc DNA3.1 group and HMGA1 group through MTT test and flat-panel cloning experiments,we found that HMGA1 can promote T47 D breast cancer cell proliferation.Conclusion: 1.HMGA1 could promote the glycolytic pathway of T47 D cells buy raising the expression of HK2?PDHK?PFKP?PFKL?LDH and PKM2 enzymes.2.HMGA1 could promote the cell reproductive capacity of T47 D cells.