胃癌间充质干细胞高表达G6PD促进肿瘤细胞迁移

Overexpression of G6PD in gastric cancer mesenchymal stem cells promotes tumor cell migration

目的探究胃癌间充质干细胞(gastric cancer mesenchymal stem cells,GCMSCs)葡萄糖-6-磷酸脱氢酶(G6PD)的表达及其在肿瘤细胞迁移中的作用。方法采用western blot检测G6PD在GCMSCs、骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMMSCs)和胃癌细胞来源外泌体(exosome)预处理BMMSCs中的表达水平。生化分析仪检测MSCs中G6PD的活性。活性氧(ROS)探针检测MSCs中ROS的表达水平。台盼蓝染色法检测细胞存活率。细胞迁移试验检测G6PD对MSCs和胃癌细胞迁移的影响。使用CCK-8试剂盒检测MSCs增殖能力。结果GCMSCs中G6PD的表达量及活性较BMMSCs明显升高(t分别为7.497、11.75,P<0.01),且GCMSCs促进胃癌细胞迁移能力较BMMSCs明显增强(P<0.05)。胃癌细胞来源的exosome通过调节P53,上调BMMSCs中G6PD的表达水平(P<0.01)。G6PD可清除MSCs细胞内活性氧,维持MSCs在低胎牛血清(t=11.29,P<0.01)或H2O2(t=3.444,P<0.05)的条件下存活,并促进MSCs增殖和迁移(P<0.01)。胃癌患者肿瘤基质细胞G6PD的表达与胃癌组织大小及转移有关(P<0.05)。结论GCMSCs通过高表达G6PD促进胃癌细胞迁移。肿瘤基质G6PD有望成为肿瘤诊断和预后监测的潜在标志物。

Objective To investigate the expression of glucose-6-phosphate dehydrogenase(G6PD)in gastric cancer mesenchymal stem cells(GCMSCs)and its role in tumor cell migration.Methods The expression levels of G6PD in GCMSCs,bone marrow mesenchymal stem cells(BMMSCs),and BMMSCs treated with exosomes derived from gastric cancer cells were detected by Western blot.The activity of G6PD in MSCs were determined by a biochemical analyzer.The levels of reactive oxygen species(ROS)in MSCs were detected by ROS probes.Cell viability was detected by the trypan blue staining method.The effects of G6PD on the migration of MSCs and gastric cancer cells were determined by the cell migration experiment.The proliferation of MSCs was detected by the cell counting kit-8(CCK-8)method.Results The expression levels and activity of G6PD in GCMSCs were significantly higher than that in BMMSCs(t=7.497 and 11.75,P<0.01),and the ability of GCMSCs promoting the migration of gastric cancer cells was significantly higher than that of BMMSCs(P<0.05).The exosomes derived from gastric cancer cells up-regulated the expression of G6PD in BMMSCs by regulating P53(P<0.01).G6PD could scavenge ROS in MSCs,maintain the survival of MSCs in the condition of low fetal bovine serum(t=11.29,P<0.01)or H2O2(t=3.444,P<0.05),and promote the proliferation and migration of MSCs(P<0.01).The expression levels of G6PD in tumor stromal cells of gastric cancer patients were related to the size and metastasis of gastric cancer(P<0.05).Conclusion GCMSCs may promote the migration of gastric cancer cells by overexpressing G6PD.G6PD in tumor stroma is expected to be a potential marker for tumor diagnosis and prognosis monitoring.