Hara A, Niikawa S, Zhang W, Sakai N, Yamada H, Yoshimi N, Mori H
Department of Neurosurgery, Gifu University School of Medicine, Japan.
J Neurooncol. 1993 Sep;17(3):205-13. doi: 10.1007/BF01049976.
Expression of the placental form of glutathione S-transferase (GST-P) in 1-(4-amino-2-methyl-5-pyrimidinyl) methyl-3-(2-chloro-ethyl)-3-nitrosourea hydrochloride (ACNU)-sensitive 9L and C6 glioma cells, and ACNU-resistant 9L (9LR) and C6 (C6R) glioma cells was investigated by Northern blot analysis for GST-P mRNA and Western blot analysis for GST-P protein. The sensitivity of 9L, 9LR, C6 and C6R cell lines to ACNU was evaluated by microculture tetrazolium assay. Localization of GSTP-P protein in these cell lines was investigated by immunocytochemical method. Expression level of GST-P mRNA in 9LR cells was 3 times that of 9L cells and the level of GST-P protein in 9LR cells was 1.7 times that of 9L cells. On the contrary, the amount of GST-P mRNA of C6R cells was 1.3-fold larger than C6 cells and that of GST-P protein of C6R cells was 1.3-fold larger than C6 cells. Immunocytochemical investigation revealed that 9LR cells had stronger expression of GST-P in their cytoplasm than 9L cells. Expression of GST-P in both C6R and C6 cells was less than 9L and 9LR cells, and the amount was similar to each other. The present study suggests that GST-P may play an important role in detoxification of anti-cancer drugs in some glioma cells.
通过对谷胱甘肽S-转移酶（GST-P）胎盘形式的mRNA进行Northern印迹分析以及对GST-P蛋白进行Western印迹分析，研究了1-(4-氨基-2-甲基-5-嘧啶基)甲基-3-(2-氯乙基)-3-亚硝基脲盐酸盐（ACNU）敏感的9L和C6胶质瘤细胞，以及ACNU耐药的9L（9LR）和C6（C6R）胶质瘤细胞中GST-P的表达情况。通过微量培养四氮唑蓝法评估9L、9LR、C6和C6R细胞系对ACNU的敏感性。采用免疫细胞化学方法研究这些细胞系中GSTP-P蛋白的定位。9LR细胞中GST-P mRNA的表达水平是9L细胞的3倍，9LR细胞中GST-P蛋白的水平是9L细胞的1.7倍。相反，C6R细胞中GST-P mRNA的量比C6细胞大1.3倍，C6R细胞中GST-P蛋白的量比C6细胞大1.3倍。免疫细胞化学研究显示，9LR细胞胞质中GST-P的表达比9L细胞更强。C6R和C6细胞中GST-P的表达均低于9L和9LR细胞，且二者表达量相似。本研究表明，GST-P可能在某些胶质瘤细胞的抗癌药物解毒过程中发挥重要作用。
