Expression of Two Types of Nitric Oxide Synthase mRNA in Human Neuroblastoma Cell Lines-猫眼课题宝

人神经母细胞瘤细胞系中两种一氧化氮合酶 mRNA 的表达

DOI：

发表时间：

1994

期刊：

Journal of Neurochemistry

影响因子：

4.7

通讯作者：

中科院分区：

医学2区

文献类型：

作者：

H. Fujisawa;T. Ogura;Y. Kurashima;Takeshi Yokoyama;Junkoh Yamashita;H. Esumi

翻译后摘要：一氧化氮合酶（NOS）的表达进行了研究，在九个人神经母细胞瘤和两个人胶质母细胞瘤细胞系。以人n-NOS cDNA为探针，通过北方印迹分析，在9个神经母细胞瘤细胞系中的4个中检测到约10 kb的神经元NOS（n-NOS）mRNA。在随后的逆转录聚合酶链反应（RT-PCR）研究中，在另一种神经母细胞瘤细胞系中也检测到n-NOS mRNA的表达，但在其他四种神经母细胞瘤细胞系或胶质母细胞瘤细胞系中未观察到n-NOS mRNA的表达。在表达n-NOS mRNA的神经母细胞瘤细胞系中，NOS活性水平与n-NOS mRNA表达水平具有良好的相关性。Western blot分析显示，神经母细胞瘤细胞中表达的n-NOS是一种与抗n-NOS抗体反应的160-kDa蛋白。通过使用RT-PCR方法，在人神经母细胞瘤细胞中检测到一个短的n-NOS（n-NOS-2）mRNA，其具有整个n-NOS（n-NOS-1）mRNA的315 bp框内缺失。首次证明了人类n-NOS mRNA的结构多样性。

Abstract: Expression of nitric oxide synthase (NOS) was studied in nine human neuroblastoma and two human glioblastoma cell lines. Neuronal NOS (n‐NOS) mRNA of ∼10 kb was detected in four of the nine neuroblastoma cell lines by northern blot analysis using human n‐NOS cDNA as a probe. Expression of the n‐NOS mRNA was also detected in another neuroblastoma cell line in a subsequent reverse transcriptase polymerase chain reaction (RT‐PCR) study, but no n‐NOS mRNA expression was observed in the other four neuroblastoma cell lines or in the glioblastoma cell lines. The level of NOS activity correlated well with that of n‐NOS mRNA expression in neuroblastoma cell lines expressing n‐NOS mRNA. Western blot analysis showed that the n‐NOS expressed in neuroblastoma cells was a 160‐kDa protein reacted with anti‐n‐NOS antibody. By using the RT‐PCR method, a short n‐NOS (n‐NOS‐2) mRNA with a 315‐bp inframe deletion from the entire n‐NOS (n‐NOS‐1) mRNA was detected in the human neuroblastoma cells. The structural diversity of human n‐NOS mRNA was demonstrated for the first time.