高糖状态对前列腺癌细胞氧化应激状态的影响

摘要

目的 探讨高糖状态对前列腺癌细胞氧化应激状态的影响.方法 人前列腺癌细胞株LNCap经过不同葡萄糖浓度处理后,采用DCFH-DA荧光探针检测LNCap细胞活性氧(ROS)水平,采用过氧化氢(H2O2)、总超氧化物歧化酶(T-SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPX)试剂盒检测LNCap细胞中H2O2水平以及各抗氧化酶活性;采用Western blot检测LNCap细胞中SOD1和SOD2蛋白表达水平;LNCap细胞转染si-SOD2后检测H2O2水平.结果 LNCap细胞内ROS和H2O2水平随葡萄糖浓度升高而升高,且呈浓度依赖性(P＜0.05);LNCap细胞内T-SOD、CAT活性在高糖环境中明显增强(P＜0.05),而GPX活性无明显变化(P＞0.05);在高糖环境中,LNCap细胞内SOD2表达水平明显升高(P＜0.05),而SOD1表达水平无明显变化(P＞0.05);SOD2基因沉默后,高糖环境下LNCap细胞内H2O2水平明显降低(P＜0.05).结论 高糖状态对前列腺癌细胞氧化应激水平具有促进作用,H2O2水平的升高可能受SOD2表达的调控.%Objective To explore the effects of high glucose state on oxidative stress in prostate cancer cells.Methods Human prostate cancer cell line LNCap was treated with different glucose concentrations,and the level of LNCap cells reactive oxygen species (ROS) was detected by DCFH-DA fluorescent probe.The hydrogen peroxide (H2O2),total superoxide dismutase (T-SOD),catalase (CAT) and glutathione peroxidase (GPX) kits were used to detect H2O2 level and antioxidant enzymes activities in LNCap cells.Western blot was used to detect the expression levels of SOD1 and SOD2 protein in LNCap cells.The level of H2O2 was detected after LNCap cells transfected with si-SOD2.Results The levels of ROS and H2O2 in LNCap cells were increased with the increase of glucose concentration,and were concentration-de pendent (P ＜ 0.05).The activities ofT-SOD and CAT in LNCap cells were increased significantly in the high glucose environment (P ＜ 0.05),but there was no significant change in the activity of GPX (P ＞ 0.05).In the high glucose environment,the expression level of SOD2 in LNCap ceils was increased significantly (P ＜ 0.05),but the expression level of SOD1 was not significantly changed (P ＞ 0.05).After the SOD2 gene was silenced,the level of H2O2 in the LNCap cells was decreased significantly in the high glucose environment (P ＜ 0.05).Conclusion High glucose state can promote the oxidative stress level of prostate cancer cells,and the increase of H2O2 level may be regulated by the expression of SOD2.