天马颗粒Ⅲ含药血清调控HK2表达对结直肠癌细胞糖酵解的影响OA

Objective To investigate the regulatory effect of Tianma Granule Ⅲ medicated serum(TMKL)on glycolysis of colorectal cancer cells and clarify the potential mechanism of hexokinase 2(HK2)in this process.Methods Using colorectal cancer SW480 cells as the research subject,the optimal concentration of TMKL for acting on SW480 was screened by the CCK-8 method.After constructing HK2-knockdown SW480 cells using plasmid transfection technology,the cells were divided into the blank transfection+blank serum group(BV+BS group),HK2 knockdown+blank serum group(sh-HK2+BS group),blank transfection+containing serum group(BV+TMKL group),and HK2 knockdown+containing serum group(sh-HK2+TMKL group).The proliferation ability of each group of cells was detected by the CCK-8 method,and glycolytic activity was assessed by measuring glucose consumption and lactate production.The expression of key glycolytic proteins glucose transporter 1(GLUT1)and pyruvate kinase M2(PKM2)were detected by Western blot and qRT-PCR.The co-expression pattern and prognostic value of HK2,GLUT1 and PKM2 in colorectal cancer were also analyzed using TCGA database.Results Compared with BV+BS group,the cell proliferation ability of sh-HK2+BS group and BV+TMKL group significantly decreased(P<0.05,P<0.001),with both glucose consumption and lactate production significantly reduced(P<0.001),the expression of HK2 protein and mRNA was significantly reduced in sh-HK2+BS group(P<0.01),both sh-HK2+BS group and BV+TMKL group exhibited significant reductions in the expression of GLUT1 and PKM2 protein and mRNA(P<0.01,P<0.001).TCGA database analysis showed that HK2 was weakly positively correlated with GLUT1 and pkm2 expression(Pearson correlation coefficients were 0.17 and 0.23,respectively,P<0.001),and low HK2 expression with high expression of GLUT1,PKM2 was associated with a shortened progression-free survival period(P<0.05).Conclusion TMKL can inhibit glycolysis in colorectal cancer cells,and its mechanism may be related to the regulation of HK2 levels,downregulating the expression of key glycolytic molecules GLUT1 and PKM2.