a Institute of Medical and Clinical Biochemistry, Faculty of Medicine , University of Belgrade , Belgrade , Serbia.
b Clinic of Urology, Clinical Centre of Serbia, Faculty of Medicine , University of Belgrade , Belgrade , Serbia.
Redox Rep. 2017 Nov;22(6):486-492. doi: 10.1080/13510002.2017.1299909. Epub 2017 Mar 13.
Newly discovered glutathione transferase omega 1 (GSTO1-1) plays an important role in the glutathionylation cycle, a significant mechanism of protein function regulation. GSTO1-1 expression pattern has not been studied in transitional cell carcinoma (TCC), as yet.
A total of 56 TCC tumor and corresponding non-tumor specimens were investigated. Glutathione content and thioltransferase activity were measured spectrophotometrically. Protein-glutathione mixed disulfides were measured fluorimetrically. GSTO1-1 expression was determined by immunoblot and qPCR. Immunoprecipitation with GSTO1-1 antibody was followed by immunoblot using anti-GSTO1, GSTP1, c-Jun, JNK, Akt, phospho-Akt, and ASK1 antibody, while for the total S-glutathionylation levels non-reducing electrophoresis was performed.
The contents of reduced glutathione and thioltransferase activity were significantly increased in tumor compared to non-tumor tissue. The increased GSTO1 expression in tumor tissue showed clear correlation with grade and stage. However, decreased total protein glutathionylation level in tumor compared to non-tumor samples was found. Immunoprecipitation has shown an association of GSTO1-1 with GSTP1, Akt, phospho-Akt, and ASK1 proteins.
GSTO1 deglutathionylase activity suggests its potential important role in redox perturbations present in TCC. Increased GSTO1-1 expression might contribute to TCC development and/or progression supporting the notion that GSTO1-1 may be a promising novel cancer target.
新发现的谷胱甘肽转移酶 ω1（GSTO1-1）在谷胱甘肽化循环中发挥重要作用，该循环是蛋白质功能调节的重要机制。目前尚未研究 GSTO1-1 在移行细胞癌（TCC）中的表达模式。
共检测了 56 例 TCC 肿瘤和相应的非肿瘤标本。使用分光光度法测量谷胱甘肽含量和硫基转移酶活性。使用荧光法测量蛋白质-谷胱甘肽混合二硫化物。通过免疫印迹和 qPCR 测定 GSTO1-1 的表达。用 GSTO1-1 抗体进行免疫沉淀，然后用 GSTO1、GSTP1、c-Jun、JNK、Akt、磷酸化 Akt 和 ASK1 抗体进行免疫印迹，而对于总 S-谷胱甘肽化水平则进行非还原电泳。
与非肿瘤组织相比，肿瘤组织中还原型谷胱甘肽的含量和硫基转移酶活性显著增加。肿瘤组织中 GSTO1 表达的增加与分级和分期有明显的相关性。然而，与非肿瘤样本相比，肿瘤样本中的总蛋白谷胱甘肽化水平降低。免疫沉淀表明 GSTO1-1 与 GSTP1、Akt、磷酸化 Akt 和 ASK1 蛋白有关。
GSTO1 脱谷胱甘肽酶活性表明其在 TCC 中存在的氧化还原扰动中具有潜在的重要作用。GSTO1-1 表达增加可能有助于 TCC 的发生和/或发展，支持 GSTO1-1 可能是一个有前途的新型癌症靶点的观点。
