Down-regulation Of The Effect Of PKM2 On Osteoblast Differentiation Phenotype In Osteosarcoma Cells

Objective: The relative expression of PKM2 in osteosarcoma MG-63 cells was studied experimentally to study the possible changes in the phenotype of THP-1 derived monocytes in the tumor microenvironment after induction into macrophages.A preliminary study of the correlation between PKM2 and tumor-associated macrophage TAM in osteosarcoma.Method:1.PKM2 levels in osteosarcoma MG-63 cells were significantly reduced by siRNA interference technology.2.Determination of PKM2 expression in MG-63 cells transfected with siRNA by qRT-PCR.3.The indirect co-culture system was constructed by Transwell chamber,and the osteosarcoma MG-63 cells with low expression of PKM2 were combined with the THP-1 cells treated with PMA to form an indirect co-culture model to simulate the tumor microenvironment.4.Using flow cytometry to detect the proportion of CD209 in mononuclear macrophages that mimic the tumor microenvironment,and to analyze the differentiated phenotype after macrophage induction according to the relative proportion.Results : 1.In order to study the expression of PKM2 in osteosarcoma cells in tumor microenvironment,the design,synthesis and transfection of siRNA showed that osteosarcoma MG-63 cells can significantly reduce the expression of PKM2 by PKM2-siRNA.Compared with the normal culture group,the change of PKM2 expression in the experimental group was significantly lower,and the difference was statistically significant(P<0.05).2.The THP-1 cells co-cultured with PKM2 osteosarcoma MG-63 in the experimental group after flow-through test showed that the percentage of CD209+ cells decreased(P<0.05 compared with the control group and the normal culture group).Conclusions:Decreased expression of PKM2 in osteosarcoma reduces the number of monocytes differentiated into M2 type macrophages.