BACKGROUND Gestational diabetes mellitus(GDM)is associated with a heightened level of oxidative stress,which is characterized by the overproduction of reactive oxygen species(ROS)from mitochondria.Previous studies sho...BACKGROUND Gestational diabetes mellitus(GDM)is associated with a heightened level of oxidative stress,which is characterized by the overproduction of reactive oxygen species(ROS)from mitochondria.Previous studies showed that mitochondrial dysfunction is regulated by dynamin-related protein 1(Drp1)and p66Shc in GDM.AIM The aim was to investigate the expression of Drp1 and p66Shc and their possible mechanisms in the pathogenesis of GDM.METHODS A total of 30 pregnant women,15 with GDM and 15 without GDM,were enrolled.Peripheral blood mononuclear cells and placental tissue were collected.The human JEG3 trophoblast cell line was cultivated in 5.5 mmol/L or 30 mmol/L glucose and transfected with wild-type(wt)-p66Shc and p66Shc siRNA.P66Shc and Drp1 mRNA levels were detected by quantitative real-time polymerase chain reaction.The expression of p66Shc and Drp1 was assayed by immunohistochemistry and western blotting.ROS was assayed by dihydroethidium staining.RESULTS The p66Shc mRNA level was increased in the serum(P<0.01)and placentas(P<0.01)of women with GDM,and the expression of Drp1 mRNA and protein were also increased in placentas(P<0.05).In JEG3 cells treated with 30 mmol/L glucose,the mRNA and protein expression of p66Shc and Drp1 were increased at 24 h(both P<0.05),48 h(both P<0.01)and 72 h(both P<0.001).ROS expression was also increased.High levels of Drp1 and ROS expression were detected in JEG3 cells transfected with wt-p66Shc(P<0.01),and low levels were detected in JEG3 cells transfected with p66Shc siRNA(P<0.05).CONCLUSION The upregulated expression of Drp1 and p66shc may contribute to the occurrence and development of GDM.Regulation of the mitochondrial fusion-fission balance could be a novel strategy for GDM treatment.展开更多
文摘目的观察p66Shc蛋白在膀胱癌中的表达,分析其表达与膀胱癌临床分期、病理类型、转移等的关系,以及p66Shc影响膀胱癌细胞增殖、凋亡的分子机制。方法 应用免疫组化法和Western blot技术检测32例膀胱癌中p66Shc蛋白的表达,分析p66Shc蛋白表达和临床病理特征的关系。Western blot技术检测T24细胞中PKCβ和p66Shc蛋白及其磷酸化水平,以及p53、Bax和BCL-2蛋白水平。结果 膀胱癌组织中p66Shc阳性率为65.6%(21/32),其在T3+T4期膀胱癌中的表达显著高于T1+T2期膀胱癌( P <0.05);p66Shc表达与患者年龄、性别、分化程度无关( P >0.05)。LY333531下调T24细胞中PKCβ和p66Shc蛋白及其磷酸化水平,以及下调p53、Bax和上调BCL-2的蛋白水平。结论 p66Shc蛋白在膀胱癌中的表达与膀胱癌临床分期相关,随着pTNM分期增加而表达增多。p66Shc磷酸化促进肿瘤细胞的增殖和凋亡。
基金The Scientific Research Fund of Qilu Hospital(Qingdao),No.QDKY2015ZD04.
文摘BACKGROUND Gestational diabetes mellitus(GDM)is associated with a heightened level of oxidative stress,which is characterized by the overproduction of reactive oxygen species(ROS)from mitochondria.Previous studies showed that mitochondrial dysfunction is regulated by dynamin-related protein 1(Drp1)and p66Shc in GDM.AIM The aim was to investigate the expression of Drp1 and p66Shc and their possible mechanisms in the pathogenesis of GDM.METHODS A total of 30 pregnant women,15 with GDM and 15 without GDM,were enrolled.Peripheral blood mononuclear cells and placental tissue were collected.The human JEG3 trophoblast cell line was cultivated in 5.5 mmol/L or 30 mmol/L glucose and transfected with wild-type(wt)-p66Shc and p66Shc siRNA.P66Shc and Drp1 mRNA levels were detected by quantitative real-time polymerase chain reaction.The expression of p66Shc and Drp1 was assayed by immunohistochemistry and western blotting.ROS was assayed by dihydroethidium staining.RESULTS The p66Shc mRNA level was increased in the serum(P<0.01)and placentas(P<0.01)of women with GDM,and the expression of Drp1 mRNA and protein were also increased in placentas(P<0.05).In JEG3 cells treated with 30 mmol/L glucose,the mRNA and protein expression of p66Shc and Drp1 were increased at 24 h(both P<0.05),48 h(both P<0.01)and 72 h(both P<0.001).ROS expression was also increased.High levels of Drp1 and ROS expression were detected in JEG3 cells transfected with wt-p66Shc(P<0.01),and low levels were detected in JEG3 cells transfected with p66Shc siRNA(P<0.05).CONCLUSION The upregulated expression of Drp1 and p66shc may contribute to the occurrence and development of GDM.Regulation of the mitochondrial fusion-fission balance could be a novel strategy for GDM treatment.
