国家十五科技攻关项目(2002BA711A08,2002BA711A03)和国家自然科学基金资助项目(30330560,30271403,30200160,30300064).
This work was supported by grants from The Special Funds of Major State Basic Research of China (2002BA711A03,2002BA711A08) and The National Natural Sciences Foundation of China (30330560,30271403, 30200160,30300064).
NGX6 是一个新克隆的鼻咽癌候选抑瘤基因 . 为进一步研究其功能,在构建 NGX6 的真核表达载体 NGX6/pcDNA3.1(+) 基础上,通过脂质体转染方法将 NGX6 基因导入鼻咽癌细胞株 SUNE-1 的亚株 5-8F 细胞 ( 具高成瘤高转移潜能 ) 中,并用 RT-PCR 和 RNA 印迹鉴定,建立了稳定表达 NGX6 基因的 5-8F 细胞系 . 借助细胞生长曲线、软琼脂集落形成实验对转染细胞的生物学行为进行了检测,同时采用包含 1 176 个与肿瘤学相关的基因 cDNA 微阵列,分析了 NGX6 基因转染对 5-8F 细胞基因表达谱的影响 . 结果显示:转染了 NGX6 基因的 5-8F 细胞的生长速度明显减慢,在软琼脂中集落形成率较对照组显著下降 (P < 0.05) ,发现 NGX6 基因的转染能够上调 5-8F 细胞中 p19 、 catenin α 2 、 desmoglein 1 等基因的表达,同时下调 EphB4 、 TIE2 、 vitronectin 等基因的表达 . 综上所述, NGX6 基因可以抑制 5-8F 细胞的恶性生物学行为,并影响一些与细胞周期、细胞黏附和血管生成有关的基因的表达 . 上述结果为鼻咽癌转移分子机制的阐明提供了重要的线索 .
NGX6 is a candidate tumor suppressor gene which was isolated by location candidate cloning strategy. In order to explore the effect of NGX6, the mammal expression vector of NGX6/pcDNA3.1(+) was transfected into 5-8F cell ( with high ability of metastasis ) by liposome. The integration of the exogenous vector DNA and the expression of NGX6 were detected by Northern blot and RT-PCR respectively. The cytobiological characterization of positive clone was analyzed by growth curves of cells and soft agar assay. cDNA array techniques were used to profile the potential targets of NGX6. The results showed that 5-8F cells with overexpression of NGX6 grew slower than that of control, and NGX6 could influence the expression of cell cycle, cell adhesion and angiopoiesis molecules, such as up-regulating p19, catenin α2, desmoglein 1,down-regulating EphB4, TIE2, vitronectin. These data revealed that NGX6 might play a role in tumor metastasis in nasopharyngeal carcinoma (NPC).
王莉莉,张秋红,马 健,彭 聪,曹 利,唐 珂,李小玲,李桂源. NGX6 基因对高转移鼻咽癌细胞株 5-8F 细胞的影响[J].生物化学与生物物理进展,2005,32(7):618-624
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