应用cDNA微阵列技术研究EB病毒LMP1介导的鼻咽癌中转移相关基因的表达

探讨EB病毒基因组编码的癌蛋白LMP1对鼻咽癌细胞中转移相关基因表达的影响.采用蛋白质印迹法检测在强力霉素(Dox)诱导下,鼻咽癌细胞系pTet-on-LMP1 HNE2(L7细胞)中LMP1表达的时效和量效关系.应用cDNA微阵列技术建立诱导性LMP1介导鼻咽癌细胞中转移相关基因差异表达谱;运用RT-PCR验证cDNA微阵列筛选差异基因表达的可靠性.与LMP1不表达的L7细胞比较,LMP1高表达的L7细胞中7个基因的表达显著上调,12个基因的表达显著下调.随机选择其中4个基因进行RT-PCR,结果显示,这些基因表达阳性,且与微阵列中的变化趋势一致.LMP1可能通过激活和/或抑制一些转移相关基因的表达而参与鼻咽癌转移过程.

Study on Expression of Metastasis-associated Genes in Nasopharyngeal Carcinoma Cell Line Mediated by EBV-LMP1 with cDNA Microarray

This paper was to investigate the effect of EB virus encoded latent membrane protein 1(LMP1) on expression of metastasis associated genes in nasopharyngeal carcinoma.Time dependent and dose dependent correlation between Dox and inducible LMP1 were confirmed with Western blotting,differential display was established to study the metastasis associated genes induced by LMP1 in nasopharyngeal carcinoma cell lines with cDNA microarray technique.Results of cDNA micrarray was confirmed by RT PCR.Compared to uninduced LMP1 negative NPC cell lines,7 genes were up regulated and 12 genes were down regulated by high expressed LMP1,changes of expression of 4 genes selected at random for RT PCR were consistant with results from cDNA microarray.It is possible for LMP1 to activate or inhibit expressions of metastasis associated genes,thus to be involvd in metastasis of nasopharyngeal carcinoma cells.