葡萄糖-6-磷酸脱氢酶对食管癌细胞侵袭能力的影响及其机制

Effect of glucose-6-phosphate dehydrogenase on invasion of esophageal cancer cells and its molecular mechanism

目的探讨葡萄糖-6-磷酸脱氢酶(G6PD)对食管癌细胞侵袭能力的影响及其机制。方法选取2017年6月至2021年6月河南省人民医院收集的54例食管癌和癌旁组织作为研究对象,采用免疫组织化学分析食管癌和癌旁组织G6PD表达水平;在人食管癌细胞株EC109细胞采用慢病毒感染建立对照组和G6PD组,采用划痕实验分析细胞迁移能力;采用Transwell分析细胞侵袭能力;采用蛋白质免疫印迹分析细胞上皮-间充质转化能力;组间计量数据比较采用t检验。结果G6PD主要表达于细胞质中,呈黄棕色。旁组织中G6PD表达水平(63.87±12.89)明显低于食管癌组织G6PD表达水平(146.56±24.38),差异有统计学意义(t=22.030,P<0.05).对照组细胞划痕愈合率[(67.62±4.99)%]明显低于G6PD组划痕愈合率[(91.21±3.57)%],差异有统计学意义(t=9.420,P<0.05)。对照组细胞黏着斑激酶(FAK)蛋白表达水平(1.01±0.08)明显低于G6PD组(2.01±0.011),差异有统计学意义(t=17.510,P<0.05)。对照组细胞迁移数量[(115.67±10.52)个]明显低于G6PD组划痕愈合率[(187.00±12.26)个],差异有统计学意义(t=10.810,P<0.05)。对照组细胞基质金属蛋白酶(MMP)-9蛋白表达水平(0.70±0.10)明显低于G6PD组(1.36±0.17),差异有统计学意义(t=17.510,P<0.05)。对照组细胞上皮标志物E-cadherin蛋白表达水平(1.06±0.05)明显高于G6PD组细胞(0.72±0.08),差异有统计学意义(t=8.724,P<0.05)。对照组细胞间质标志物波形蛋白(Vimentin)和N-钙黏蛋白(N-cadherin)表达水平(1.09±0.09、0.83±0.05)明显低于G6PD组细胞(1.96±0.21、1.50±0.12),差异有统计学意义(t=9.274、13.170,P<0.05)。结论G6PD在食管癌组织呈高表达,并参与食管癌细胞的迁移、侵袭和上皮-间充质转化等过程。

Objective To investigate the effect of glucose-6-phosphate dehydrogenase(G6PD)on the invasion of esophageal cancer cells and its molecular mechanism.Methods Totally,54 cases of esophageal cancer and adjacent tissues collected in our hospital from June 2017 to June 2021 were selected as the study objects,and the expression level of G6PD in esophageal cancer and adjacent tissues was analyzed by immunohistochemistry.The control group and G6PD group were established by lentivirus infection in human esophageal cancer cell line EC109,and the cell migration ability was analyzed by scratch test.The invasiveness of cells was analyzed by Transwell.The ability of epithelial mesenchymal transformation was analyzed by Western blotting.The measurement data between groups were compared by t test.Results G6PD was mainly expressed in the cytoplasm,which was yellowish brown.The expression level of G6PD in adjacent tissues(63.87±12.89)was significantly lower than that in esophageal cancer tissues(146.56±24.38,t=22.030,P<0.05).The scratch healing rate of control group[(67.62±4.99)%]was significantly lower than that of G6PD group[(91.21±3.57)%,t=9.420,P<0.05].The expression level of focal adhesion kinase(FAK)protein in control group was significantly lower than in the G6PD group(1.01±0.08 vs.2.01±0.011,t=17.510,P<0.05).The number of migrating cells[(115.67±10.52)]in the control group was significantly less than in the G6PD group[(187.00±12.26),t=10.810,P<0.05].The expression level of MMP-9 protein in the control group(0.70±0.10)was significantly lower than that in the G6PD group(1.36±0.17,t=17.510,P<0.05).The expression level of E-cadherin protein in the control group(1.06±0.05)was significantly higher than that in the G6PD group(0.72±0.08,t=8.724,P<0.05).The expression levels of Vimentin and N-cadherin protein in the control group(1.09±0.09 and 0.83±0.05)were significantly higher than those in G6PD group(1.96±0.21 and 1.50±0.12,t=9.274,13.170,P<0.05).Conclusion G6PD is highly expressed in esophageal cancer tissues and participates in the migration,invasion and epithelial mesenchymal transformation of esophageal cancer cells.