高糖环境对卵巢癌细胞侵袭和迁移的影响及其机制实验研究

Effect of high glucose environment on the invasion and migration of ovarian cancer cells and its mechanism

目的:探究高糖环境对卵巢癌细胞侵袭和迁移的影响及其可能的作用机制。方法:将人卵巢癌细胞系SKOV-3分为正常糖组(低糖型培养基培养)、高糖组(高糖型培养基培养)、高糖+过氧化氢酶(CAT)组(高糖型培养基+400 U/ml CAT培养)、高糖+丝裂原活化的细胞外信号调节激酶(MEK)抑制剂PD98059组(高糖型培养基+50μmol/ml PD98059培养)。采用过氧化氢(H_(2)O_(2))检测试剂盒测定细胞内H_(2)O_(2)水平。划痕实验检测细胞迁移能力。Transwell实验检测细胞侵袭能力。免疫印迹法检测细胞E钙黏蛋白(E-cadherin)、N钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、细胞外信号调节激酶(ERK)、p-ERK、核因子-κB(NF-κB)、p-NF-κB蛋白表达。RT-qPCR检测细胞E-cadherin、N-cadherin、Vimentin mRNA表达。结果:与正常糖组比较,高糖组H_(2)O_(2)水平、24 h细胞迁移率、侵袭细胞数、p-ERK、p-NF-κB蛋白表达水平和N-cadherin、Vimentin蛋白及mRNA表达水平升高,E-cadherin蛋白及mRNA表达水平降低(均P<0.01)。与高糖组比较,高糖+CAT组H_(2)O_(2)水平降低,高糖+CAT组和高糖+PD98059组24 h细胞迁移率、侵袭细胞数、p-ERK及p-NF-κB蛋白表达水平和N-cadherin、Vimentin蛋白及mRNA表达水平降低,E-cadherin蛋白及mRNA表达水平升高(均P<0.01)。结论:高糖能够诱导卵巢癌细胞产生H_(2)O_(2)并激活下游ERK/NF-κB信号通路,促进卵巢癌细胞的侵袭和迁移,并影响卵巢癌细胞上皮-间充质转化(EMT)相关因子表达。

Objective:To investigate the effect of high glucose environment on invasion and migration of ovarian cancer cells and its possible mechanism.Methods:Human ovarian cancer cell line SKOV-3 was divided into normal glucose group(low glucose medium culture),high glucose group(high glucose medium culture),high glucose+catalase(CAT)group(high glucose medium+400 U/ml CAT culture),and high glucose+mitogen-actived extracellular signal-regulated kinase(MEK)inhibitor PD98059 group(high glucose medium+50μmol/ml PD98059 culture).Intracellular hydrogen peroxide(H_(2)O_(2))levels were measured by H_(2)O_(2) assay kit.Cell migration ability was detected by scratch test.Transwell assay was used to detect cell invasion ability.The expression of E-cadherin,N-cadherin,Vimentin,extracellular signal-regulated kinase(ERK),p-ERK,nuclear factor-κB(NF-κB)and p-NF-κB proteins was detected by Western blot.The expression of E-cadherin,N-cadherin and Vimentin mRNA was detected by RT-qPCR.Results:Compared with normal glucose group,H_(2)O_(2) level,24 h cell migration rate,number of invading cells,the expression levels of p-ERK,p-NF-κB proteins and the protein and mRNA expression levels of N-cadherin,Vimentin were increased,and the protein and mRNA expression levels of E-cadherin were decreased in high glucose group(all P<0.01).Compared with the high glucose group,H_(2)O_(2) levels were decreased in the high glucose+CAT group,24 h cell migration rate,number of invading cells,the expression levels of p-ERK,p-NF-κB proteins and the protein and mRNA expression levels of N-cadherin,Vimentin were decreased,the protein and mRNA expression levels of E-cadherin were increased in the high glucose+CAT and high glucose+PD98059 groups(all P<0.01).Conclusion:High glucose can induce H_(2)O_(2) production and activate downstream ERK/NF-κB signaling pathway in ovarian cancer cells,promote invasion and migration of ovarian cancer cells,and affect the expression of epithelial-mesenchymal transition(EMT)-related factors in ovarian cancer cells.