鼻咽癌肺转移相关基因的检测及筛选

Detection and screen of pulmonary metastasis-related signature genes in nasopharyngeal carcinoma

目的 通过建立鼻咽癌（nasopharyngeal carcinoma,NPC）的肺转移动物模型,运用基因芯片技术检测及筛选肺转移最相关的10种差异基因.方法 采用鼻咽癌细胞株CNE,构建稳定表达荧光酶基因的CNE/Luc稳转细胞株,尾静脉注射CNE/Luc悬液到裸鼠中,利用动物活体成像系统筛选得到肺转移裸鼠,分离肿瘤细胞,进行2次尾静脉注射筛选得到肺部转移的肿瘤细胞,运用荧光成像验证其组织特异性转移后,通过基因表达谱芯片技术,获得鼻咽癌肺转移相关的差异基因.通过文献检索筛选出相关性最高的10种基因进行分析.结果 本实验利用基因芯片技术,构建鼻咽癌原代细胞株CNE/Luc与鼻咽癌肺细胞转移相关亚群CNE/Luc-2的基因表达谱,通过软件分析,筛选出10个可能与鼻咽癌肺转移关系密切的基因,分别是TP53、PIK3CA、MET、KRAS、VEGFA、EDNRB、GSK3B、FOXO3、SOD2、BIRC3.结论 通过构建动物模型及基因芯片技术得出的10个基因对于鼻咽癌的肺转移有重要的调节作用,对下一步验证鼻咽癌肺转移相关通路有重要意义.

Objective To establish an animal model of pulmonary metastasis from nasopharyngeal carcinoma （NPC） and to investigate differential genes associated with pulmonary metastasis.Methods CNE cell line was used to construct the stable metastasis CNE/Luc cell line which steadily expresses the fluorescent enzyme genes.The CNE/Luc cells were injected into immunodeficiency mice through tail vein,and with the in vivo imaging technology,the mice with pulmonary metastasis were filtered out.The pulmonary metastasis cells,were separated and injected into the tail vein of other nude mice to obtain the tissue-specificity metastasis cells confirmed by fluorescent imaging system.Based on the gene chip,the differential genes associated with pulmonary metastasis for NPC were found.Results The gene expression profiles of nasopharyngeal carcinoma cell line CNE/Luc and their lung metastasis-associated subpopulation CNE/Luc-2 were constructed by gene chip technology.Ten metastasis-related genes were screened by software analysis,namely as TP53,PIK3CA,MET,KRAS,VEGFA,EDNRB,GSK3B,FOXO3,SOD2,and BIRC3.Conclusions Some genes including TP53,PIK3CA,MET,KRAS,VEGFA,EDNRB,GSK3B,FOXO3,SOD2,and BIRC3 are indicated to have important roles in the lung metastasis of nasopharyngeal carcinoma.