Hexokinase and glucose-6-phosphatase activity in woodchuck model of hepatitis virus-induced hepatocellular carcinoma
Hexokinase and glucose-6-phosphatase activity in woodchuck model of hepatitis virus-induced hepatocellular carcinoma
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DOI：
10.1016/j.cbpc.2006.02.005
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发表时间：
2006-06-01
影响因子：
3.9
通讯作者：
Lee, Zhenghong
中科院分区：
文献类型：
--
作者：
Kuang, Yu;Schomisch, Steve J.;Lee, Zhenghong
2-脱氧-2-[F-18]氟- d -葡萄糖（[F-18] FDG）用于土拨鼠（Mannota monax）肝细胞癌（HCC）模型的PET成像。FDG在该动物模型上的有效性需要根据假设的机制进行验证。本研究检测了土拨鼠肝细胞癌模型中参与葡萄糖或[F-18] FDG代谢的两种关键酶己糖激酶（HK）和葡萄糖-6-磷酸酶（G6Pase）的酶活性，这在以前的研究中没有发现。动态PET扫描后，收获土拨鼠肝组织样本，离心匀浆。上清液用于HK活性测定，微粒体微球用于G6Pase测定。HK和G6Pase活性分别通过动力学和终点法比色反应测定。用Bradford法测定总蛋白含量，并将所有酶活性归一化。土拨鼠HCC中的HK和G6Pase活性将用于与体内PET成像数据相关联。与年龄匹配的健康土拨鼠相比，HCC模型肝脏HK水平显著升高（7.96 +/- 1.27 vs. 2.74 +/- 0.66 mU/mg蛋白，P < 0.01）， G6Pase水平显著降低（40.35 +/- 19.28 vs. 237.01 +/- 17.32 mU/mg蛋白，P < 0.01），反映糖酵解增加。此外，在同一鼠肝模型中，肝癌肝区（HK: 7.96 1.27 mU/mg蛋白；G6Pase: 40.35 +/- 19.28 mU/mg蛋白）与周围正常肝区（HK: 2.98 +/- 0.92 mU/mg蛋白；G6Pase: 140.87 +/- 30.62 mU/mg蛋白）之间HK和G6Pase活性差异显著（P < 0.01）。我们的研究表明，与正常的土拨鼠肝脏相比，肝细胞肝癌模型土拨鼠肝脏中HK活性增加，G6Pase活性降低。(c) 2006爱思唯尔公司版权所有。
2-Deoxy-2-[F-18]fluoro-D-glucose ([F-18] FDG) is used for PET imaging of woodchuck (Mannota monax) model of hepatocellular carcinoma (HCC). The usefulness of FDG on this animal model needs to be validated according to the hypothesized mechanisms. In this study, two key enzymes involved in glucose or [F-18] FDG metabolism, hexokinase (HK) and glucose-6-phophatase (G6Pase), were examined for their enzymatic activities in the woodchuck models of HCC, which has not been studied before. After dynamic PET scans, woodchuck liver tissue samples were harvested and the homogenate was centrifuged. The supernatant was used for HK activity assay and the microsomal pellet was used for G6Pase assay. HK and G6Pase activities were measured by means of colorimetric reactions via kinetic and end-point assays, respectively. Total protein content was measured by the Bradford method and used to normalize all enzyme activities. HK and G6Pase activities in woodchuck HCC will be used to correlate with in vivo PET imaging data. The woodchuck model of HCC had significantly increased levels of HK in the livers compared to the age-matching healthy woodchuck (7.96 +/- 1.27 vs. 2.74 +/- 0.66 mU/mg protein, P < 0.01) and significantly decreased levels of G6Pase compared to healthy woodchuck (40.35 +/- 19.28 vs. 237.01 +/- 17.32 mU/mg protein, P < 0.01), reflecting an increase in glycolysis. In addition, significant differences were found in HK and G6Pase activities between HCC liver region (HK: 7.96 1.27 mU/mg protein; G6Pase: 40.35 +/- 19.28 mU/mg protein) and surrounding normal liver region (HK: 2.98 +/- 0.92 mU/mg protein; G6Pase: 140.87 +/- 30.62 mU/mg protein) in the same woodchuck model of HCC (P < 0.01). Our study demonstrated an increased HK activity and a decreased G6Pase activity in liver of the woodchuck models of HCC as compared to normal woodchuck liver. (c) 2006 Elsevier Inc. All rights reserved.