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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 newly cloned nasopharyngeal cancer candidate tumor suppressor gene.To further study its function, we constructed NGX6 eukaryotic expression vector NGX6 / pcDNA3.1 (+) on the basis of liposome transfection method to NGX6 gene into The sub-line 5-8F cells (with high tumorigenicity and high metastatic potential) of nasopharyngeal carcinoma cell line SUNE-1 were identified by RT-PCR and Northern blotting to establish 5-8F cell lines stably expressing NGX6 gene.Using the help of cells The growth curve and soft agar colony formation assay were used to detect the biological behavior of the transfected cells. The cDNA microarray containing 1176 oncogenes was used to analyze the effect of NGX6 gene transfection on the gene expression profile of 5-8F cells The results showed that the growth of 5-8F cells transfected with NGX6 gene was significantly slowed down, the colony formation rate in soft agar was significantly decreased compared with the control group (P <0.05), and found that NGX6 gene transfected 5- 8F cells, downregulate the expression of EphB4, TIE2, vitronectin and other genes.In conclusion, NGX6 gene can inhibit the malignant biological behavior of 5-8F cells and affect some of the cell cycle , Cell adhesion and angiogenesis have Expression of the gene. These results provide important clues for elucidating the molecular mechanism of metastasis of nasopharyngeal carcinoma.