Study Of TNFAIP8 High Expression Promotes Prostate Cancer Cell PC3 And LNCaP Proliferation And Its Mechanism

Objective To observe the effect of TNFAIP8 gene high expression on the proliferation of prostate cancer cells PC3 and LNCaP and its possible mechanism by onstructing TNFAIP8 gene overexpression and silencing models.Methods Normal prostate epithelial cells RWPE1,prostate cells PC3,and LNCaP were selected as experimental subjects.MTT experiments were used to observe the cell proliferation ability.ELISA kits were used to detect glucose consumption and ATP production.qPCR and Western Blotting were used to detect the mRNA and protein expression of TNFAIP8 gene.The liposome transfection technology was used to construct a TNFAIP8 gene overexpression and silencing model,which was divided into an empty vector transfection group and a TNFAIP8 gene overexpression group,a control group and a TNFAIP8 gene silencing group,and observed cell proliferation,detected glucose consumption and ATP production,Seahorse XFp bioenergy analyzer detects changes in mitochondrial respiratory function,qPCR and Western Blotting detect changes in glucose metabolism-related enzymes HK1,Glut1,G6 PD,AMPK,LDHA.Results(1)Compared with prostate epithelial cells RWPE1,prostate cancer cells PC3 and LNCaP have stronger proliferation ability,PC3 cells have higher glucose consumption and LNCaP cells have higher ATP production ability.Both the mRNA expression of endogenous TNFAIP8 gene in PC3 and LNCaP cells were low,and the expression of endogenous TNFAIP8 gene protein in LNCaP cells was low.(2)Overexpression of TNFAIP8 gene,compared with the empty vector group,all three cells have enhanced proliferation capacity,increased glucose consumption,and increased ATP production.Respiratory function of mitochondria was enhanced in PC3 and LNCaP cells overexpression group.The mRNA expression of glucose metabolism enzymes HK1,Glut1,AMPK,and LDHA in RWPE1 cells increased compared with the empty vector group;HK1,Glut1,and AMPK protein expression increased compared to the empty vector group,and G6 PD was not expressed.The mRNA expression of the enzyme HK1,Glut1,and LDHA in the overexpression group of PC3 cells was increased compared with the empty vector group;Glut1,G6 PD,and AMPK protein expression were increased.LNCaP cells overexpressed AMPK and LDHA mRNA expression compared with the empty vector group;Glut1 and AMPK protein expression decreased,G6 PD was not expressed.(3)Silencing expression of TNFAIP8 gene,compared with control group,glucose consumption of RWPE1 cells,PC3 cells and LNCaP cells was reduced.PC3 cells and LNCaP cells reduced ATP production.The mitochondrial respiratory function of prostate cancer cells was lower than that of the negative control transfection group.The mRNA expressions of glucose metabolizing enzymes HK1,Glut1,G6 PD,AMPK,and LDHA were all lower than those of the control group.The expression of HK1 and AMPK protein in TNFAIP8 gene silencing group of PC3 cells and LNCaP cells decreased..Conclusion(1)TNFAIP8 gene overexpression promotes the proliferation of prostate cancer cells PC3 and LNCaP.(2)TNFAIP8 gene overexpression enhances glucose consumption,ATP production,and mitochondrial respiratory function in prostate cancer cells PC3 and LNCaP.(3)TNFAIP8 gene enhances the glucose metabolism of prostate cancer cells PC3 and LNCaP by regulating the expression of glucose metabolizing enzymes,which may be the main reason that TNFAIP8 gene overexpression promotes the proliferation of prostate cancer cells PC3 and LNCaP.