摘要
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1176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、cateninα2、desmoglein1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 p 19, catenin alpha 2, desmoglein l,down-regulating EphB4, TIIE2, vitronectin. These data revealed that NGX6 might play a role in tumor metastasis in nasopharyngeal carcinoma (NPC).
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2005年第7期618-624,共7页
Progress In Biochemistry and Biophysics
基金
国家十五科技攻关项目(2002BA711A08
2002BA711A03)
国家自然科学基金资助项目(30330560
30271403
30200160
30300064)~~
