Objective To investigate the effect of glutaminase 2(GLS2)on the proliferation and invasion ability of intrahepatic cholangiocarcinoma cells and its mechanism.Methods Human intrahepatic cholangiocarcinoma RBE cells in the logarithmic phase were randomly divided into a blank control group(Con group),a GLS2 overexpression negative control group(pcDNA-NC group),a pcDNA-GLS2 group,and a pcDNA-GLS2+phosphorylation Janus kinase(p-JAK)group.RBE cells in the Con group did not transfect any plasmid;RBE cells in the pcDNA-NC group were transfected with pcDNA-NC plasmid;RBE cells in the pcDNA-GLS2 group were transfected with pcDNA-GLS2 plasmid;RBE cells in the pcDNA-GLS2+p-JAK group were transfected with pcDNA-GLS2 plasmid and co-cultured with p-JAK peptide.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression of GLS2 mRNA in each group of cells;cell counting kit-8 was used to detect the proliferation ability of cells in each group;flow cytometry was used to detect the apoptotic ability of cells in each group;Transwell assay was used to detect the invasion ability of cells in each group;Western blot was used to detect the relative expression levels of Janus kinase 2(JAK2),phosphorylation JAK2(p-JAK2),signal transducer and activator of transcription 3(STAT3),and phosphorylation STAT3(p-STAT3)proteins in cells.Results The relative expression level of GLS2 mRNA in the pcDNA-GLS2 group was significantly higher than that in the Con group and pcDNA-NC group(P＜0.05);the relative expression level of GLS2 mRNA in the pcDNA-GLS2+p-JAK group was significantly lower than that in the pcDNA-GLS2 group(P＜0.05).The survival rate of cells in the pcDNA-GLS2 group was significantly lower than that in the Con group(P＜0.05),and compared with the pcDNA-GLS2 group,the survival rate of cells in the pcDNA-GLS2+p-JAK group was significantly increased(P＜0.05).Compared with the Con group,the apoptosis rate of cells in the pcDNA-GLS2 group significantly increased(P＜0.05);the apoptosis rate of cells in the pcDNA-GLS2+p-JAK group was significantly lower than that in the pcDNA-GLS2 group(P＜0.05).The migration number of cells in the pcDNA-GLS2 group was significantly lower than that in the Con group(P＜0.05);compared with the pcDNA-GLS2 group,the pcDNA-GLS2+p-JAK group showed a significant increase in the cell migration number(P＜0.05).Compared with the Con group,the p-JAK2/JAK2 and p-STAT3/STAT3 ratios in the pcDNA-GLS2 group were significantly reduced(P＜0.05);the p-JAK2/JAK2 and p-STAT3/STAT3 ratios in the pcDNA-GLS2+p-JAK group were significantly higher than those in the pcDNA-GLS2 group(P＜0.05).Conclusion Overexpression of GLS2 can regulate the biological activity of intrahepatic cholangiocarcinoma cells by inhibiting the JAK2/STAT3 signaling pathway,inhibit the proliferation,invasion and migration of intrahepatic cholangiocarcinoma RBE cells,and induce the cell apoptosis.
glutaminase 2Janus kinase 2/signal transducer and activator of transcription 3intrahepatic cholangiocar-cinomacell proliferationcell invasion
维普
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