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γ-谷氨酰转肽酶在转化的大鼠脑内皮细胞系中的表达

Expression of gamma-glutamyltranspeptidase in a transformed rat cerebral endothelial cell line.

作者信息

Caspers M L, Diglio C A

出版信息

Biochim Biophys Acta. 1984 Feb 17;803(1-2):1-6. doi: 10.1016/0167-4889(84)90047-8.

Abstract

Brain capillary endothelium in vivo contains high levels of gamma-glutamyltranspeptidase activity. In addition, the presence of this enzyme has been used as a marker of neoplastic cells. Normal rat cerebral endothelial cells in culture exhibit a specific activity for gamma-glutamyltranspeptidase of 2 units/10(6) cells. In vitro transformation of these cells is achieved by the use of an avian retrovirus, Schmidt-Ruppin RSV-strain D. The resultant cell line, designated RCE-T1, demonstrates a significant increase in gamma-glutamyltranspeptidase activity up to 20 units/10(6) cells in early passage levels (9-26) after which enzyme activity declines and returns to normal levels by passage 80. This variation in enzyme activity correlates with histochemical staining for this enzyme. Furthermore, the enzyme activity increases linearly over a 1000-fold range of cell concentrations. Various culture modifications do not influence this pattern of enzyme expression. These parameters include trypsin dissociation, cell freezing, degree of confluency and culture maintenance with serum or with conditioned medium obtained from passage levels exhibiting high or low enzyme activity. RCE-T1 cells will provide a unique model system to study the distribution and regulation of this enzyme during differentiation and viral carcinogenesis.

摘要

体内脑毛细血管内皮含有高水平的γ-谷氨酰转肽酶活性。此外,这种酶的存在已被用作肿瘤细胞的标志物。培养的正常大鼠脑内皮细胞对γ-谷氨酰转肽酶的比活性为2单位/10⁶个细胞。这些细胞的体外转化通过使用禽逆转录病毒施密特-鲁平劳斯肉瘤病毒D株来实现。所得的细胞系命名为RCE-T1,在传代早期水平(9-26)时γ-谷氨酰转肽酶活性显著增加,高达20单位/10⁶个细胞,此后酶活性下降,到第80代时恢复到正常水平。这种酶活性的变化与该酶的组织化学染色相关。此外,在1000倍的细胞浓度范围内,酶活性呈线性增加。各种培养条件的改变并不影响这种酶表达模式。这些参数包括胰蛋白酶解离、细胞冻存、汇合度以及用血清或从表现出高或低酶活性的传代水平获得的条件培养基进行培养维持。RCE-T1细胞将提供一个独特的模型系统,用于研究该酶在分化和病毒致癌过程中的分布和调节。

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