Effects Of HMGA1 On Glycolytic Pathways Of MCF-7 Cells

Objective: To investigate the effects of HMGA1 on the glycolytic pathway in human breast cancer cell line MCF7,and to clarify the role and molecular mechanism of HMGA1 gene in the process of glucose metabolism in breast cancer.Methods: 1.MCF 7 cell line were cultured in vitro.HMGA1 plasmid and empty plasmid were transfected respectively into MCF7 using Lipofectamine.The stable transfected cells were screened by the G418(500ug/ml).Transfection efficiency was detected by WB technique.Then the effects of HMGA1 on the proliferation of human breast cancer cell line MCF7 was detected by MTT assay and cell clone formation assay.2.By comparing the difference of glucose consumption,lactate accumulation and ATP production between MCF-7-HMGA1 and negative control group to investigate its regulation of glucose metabolism in breast cancer cells.3.The effects of HMGA1 on the key enzymes of glycolysis pathway was analyzed by protein immunoblotting in MCF-7 cells.4.The activities of the key enzymes(PFK,HK,PK,LDH)of glycolysis were further studied and the downstream target and mechanism of HMGA1 were discussed in the glycolysis pathway.Results: 1.MTT assay and cell clone formation experiments showed that: HMGA1 stably transfected human breast cancer MCF7 cells can promote the proliferation of breast cancer cells(P < 0.05).2.HMGA1 and empty vector were transfected into human breast cancer cells MCF7.The results showed that compared with pcDNA3.1group(empty vector group),the glucose consumption,lactic acid production,ATP production in HMGA1 overexpressed cells was significantly increased,and all were statistically significant(P <0.05).3.The expression of HK2,PKM2,PFKP and LDHA in the cells of HMGA1 overexpression group were significantly increased,the expression of PDH was decreased.The difference was statistically significant(P <0.05).However,there was no significant difference in the expression of HK1,PKM 1/2,PFKL and GLUT1,which was not statistically significant(P>0.05).4.The activities of HK,LDH,FPK and PK in MCF-7-HMGA1 was higher than pcDNA3.1 group,and the difference was statistically significant(P <0.05).Conclusion: 1.HMGA1 promotes MCF7 cell proliferation and clonogenic formation.2.HMGA1 can promote the glycolysis of human breast cancer MCF7 cells.