Abstract:
Reactive oxygen species (ROS) play an important and controversial role in carcinogenesis. Microsomal redox chains containing NADH- and NADPH-dependent oxidoreductases are among the main sites of intracellular ROS synthesis, but their role in the oxidative balance has not been fully studied. Here, we studied the activity of cytochrome b5 reductase (CYB5R) and cytochrome P450 reductase (CYPOR) in ovarian cancer tissues and cells isolated from peritoneal fluid, along with the antioxidant capacity of peritoneal fluid. We used the developed a chemiluminescence assay based on stimulation with NADH and NADPH, which reflects the activity of CYB5R and CYPOR, respectively. The activity of CYB5R and CYPOR was significantly higher in moderately and poorly differentiated ovarian adenocarcinomas compared with well-differentiated adenocarcinomas and cystadenomas. For the chemotherapy-resistant tumors, the activity of tissue CYB5R and CYPOR was lower compared to the non-resistant tumors. In the peritoneal fluid, the antioxidant capacity significantly increased in this series, benign tumors < well-differentiated < moderately and poorly differentiated adenocarcinomas, so the antioxidant excess was observed for moderately and poorly differentiated adenocarcinomas. The antioxidant capacity of peritoneal fluid and the activity of CYB5R and CYPOR of cells isolated from peritoneal fluid were characterized by a direct moderate correlation for moderately and poorly differentiated adenocarcinomas. These results indicate the significant role of NAD(P)H oxidoreductases and the antioxidant potential of peritoneal fluid in cancer biochemistry. The parameters studied are useful for diagnostics and prognostics. The developed assay can be used to analyze CYB5R and CYPOR activity in other tissues and cells.
摘要:
活性氧(ROS)在致癌作用中起着重要且有争议的作用。含有NADH和NADPH依赖性氧化还原酶的微粒体氧化还原链是细胞内ROS合成的主要位点,但它们在氧化平衡中的作用尚未得到充分研究。这里,我们研究了卵巢癌组织和从腹膜液中分离的细胞中细胞色素b5还原酶(CYB5R)和细胞色素P450还原酶(CYPOR)的活性,以及腹膜液的抗氧化能力。我们使用了基于NADH和NADPH刺激的化学发光测定法,这反映了CYB5R和CYPOR的活性,分别。与高分化腺癌和囊腺瘤相比,中分化和低分化卵巢腺癌的CYB5R和CYPOR活性明显更高。对于化疗耐药的肿瘤,与非耐药肿瘤相比,组织CYB5R和CYPOR的活性较低。在腹膜液中,抗氧化能力在这个系列中显著增加,良性肿瘤<高分化<中度和低分化腺癌,因此,观察到中度和低分化腺癌的抗氧化剂过量。腹膜液的抗氧化能力以及从腹膜液中分离的细胞的CYB5R和CYPOR活性的特征是与中度和低分化腺癌直接相关。这些结果表明NAD(P)H氧化还原酶和腹膜液的抗氧化潜力在癌症生物化学中的重要作用。所研究的参数对于诊断和预测是有用的。开发的测定法可用于分析其他组织和细胞中的CYB5R和CYPOR活性。