Effect of M2 pyruvate kinase(PKM2)expression in tumor associated macrophages(TAMs)on malignant phenotype of esophageal squamous cell carcinoma(ESCC)
Objective To investigate the effect of M2 pyruvate kinase(PKM2)expression in tumor associ-ated macrophages(TAMs)on the malignant phenotype of esophageal squamous cell carcinoma(ESCC)cell migration and invasion.Methods Transfection of PKM2 knockdown/overexpression plasmid with lentivirus into THP-1 cells,inducing THP-1 cells to differentiate into TAMs,which were randomly divided into PKM2 knockdown group(PKM2 KD group),PKM2 knockdown control group(PKM2 KD NC group),PKM2 overexpression group(PKM2 OE group)and PKM2 overexpression control group(PKM2 OE NC group).The expression of PKM2 and macrophage phenotype markers were detected by qRT-PCR.After cocultured with KYSE150 cells,transwell assay was used to detect the migration and invasion ability of ESCC cells.Cocultured cells were inoculated subcutaneously into nude mice to construct a nude mouse tumor bearing model.IHC was used to detect the expression of PKM2 and CD163,and ELISA was used to detect glucose and lactate levels.Results Compared with PKM2 KD NC group,the expression level of PKM2 was significantly reduced in TAMs cells in PKM2 KD group,while the expression of NOS2 was up-regulated(all P<0.05),the expression of CD163 was slightly downregulated(P>0.05),and the expres-sion of CCL5 was downregulated(P<0.05);Compared with PKM2 OE NC group,the expression level of PKM2 was significantly increased in TAMs cells in PKM2 OE group,while the expression of NOS2 was downregulated(all P<0.05),the expression of CD163 was slightly upregulated(P>0.05),and the ex-pression of CCL5 was upregulated(P<0.05);Compared with PKM2 KD NC group,the ability of migra-tion and invasion of KYSE150 cells after co-culture were significantly reduced in PKM2 KD group(all P<0.001).Compared with PKM2 OE NC group,the ability of migration and invasion of KYSE150 cells after co-culture were significantly enhanced in PKM2 OE group(all P<0.01).Compared with CO-PKM2 KD NC group,CO-PKM2 KD group showed a decrease in the volume and weight of subcutaneous transplanted tumors in nude mice with mixed cell inoculation(P>0.05).Meanwhile,the expression of PKM2 and CD163 was decreased in tumor tissue(P>0.05),and the concentration level of glucose and lactate in ser-um was significant decreased(all P<0.05).Compared with CO-PKM2 OE NC group,CO-PKM2 OE group showed an increase in the volume and weight of subcutaneous transplanted tumors in nude mice(P>0.05).The expression of PKM2 and CD163 was upregulated in tumor tissue(all P>0.05),and serum glucose and lactate levels were significantly elevated(all P<0.05).Conclusion Variation in PKM2 ex-pression affect the phenotypic differentiation of TAMs cells,and PKM2 expression in TAMs cells may af-fect the malignant phenotype of ESCC cells through anaerobic glycolysis.