Effect Of PRMT2 And PRMT2β On Glycolytic Pathways Of MCF-7 Cells

Objective: To explore the effect of PRMT2 and PRMT2β on MCF-7cells glycolysis metabolic pathways and its possible mechanism, in order to provide new research visions for breast cancers.Methods: To study the function of PRMT2 and PRMT2β in glycolytic pathways, MCF-7-PRMT2 stable cells and MCF-7-PRMT2β stable cells were used in the experiments. The experimental group was PRMT2 and PRMT2β overexpression group(over expression, OE)which had been treated with DOX(doxycyline) for 48 h, and the DOX untreated group(negative control, NC) was used as negative control. Our experiments have three parts: 1.By comparing the difference of consumed glucose,amount of produced lactic acid and level of ATP of MCF-7 cells between-DOX group and +DOX group, the regulatory effect on glucose pathway of breast cancer cells can be premilarily determined. 2. The effect of overexpression PRMT2 and PRMT2β in MCF-7 cells on key enzyme such as HK2、PK2、PFKP、LDHA、PDH and so on in glycolytic pathway can be analyzed from protein expression level by Western Blotting. 3.Through furtherly detecting the activity of glycolytic key enzyme such as HK、PK and LDH, we had preliminarily discussed the possible regulatory downstream targets and mechanisms of PRMT2 and PRMT2β in glycolytic pathway.Results: 1. The result showed that after overexpressing PRMT2 and PRMT2β, the glucose consumption level, amount of produced lactic acid and level of ATP in MCF-7 was reduced which had statistical significance(P<0.05).2. The result of Western Blot showed that, the expression level of HK2、PKM2、PFKP、PFKL、LDH in OE group was reduced and the PDH was enhanced, therefore the difference had statistical significance(P<0.05). As the expression level of HK1 、 PKM1/2 was unchanged obviously, it had no statistical significance(P>0.05).3. Through detecting the acticity of glycolytic key enzyme,we found that,compared with the NC group, the activity of HK, PK,LDH were significantly decreased in the OE group which had statistical significance(P < 0.05).Conclusion: PRMT2 and its spliceosome PRMT2β could inhibit the glycolytic pathway of MCF-7 cells.