中文摘要：

目的：研究不含POU结构域的八聚体结合蛋白（non-POU domain containing octamer-binding protein,NONO）基因沉默对鼻咽癌细胞增殖、迁移和侵袭的影响,以期探讨NONO在鼻咽癌进展中的作用。方法 ：采用蛋白质印迹法检测正常鼻咽上皮细胞NP-69以及鼻咽癌细胞CNE1和CNE2中NONO蛋白的表达。用含有靶向人NONO基因的sh RNA重组慢病毒感染鼻咽癌CNE2细胞,嘌呤霉素筛选后,采用蛋白质印迹法检测NONO基因的表达沉默效果。采用CCK-8法、平板克隆形成实验、细胞划痕愈合实验、体外Transwell迁移和侵袭实验分别检测NONO基因沉默对CNE2细胞增殖、克隆形成、迁移和侵袭能力的影响。结果：鼻咽癌细胞CNE1和CNE2中NONO蛋白表达水平比正常鼻咽上皮细胞NP-69明显升高（P值均〈0.05）。感染NONO-sh RNA重组慢病毒后,CNE2细胞中NONO基因沉默效率接近100%。与未感染或感染阴性对照慢病毒的CNE2细胞相比,NONO基因沉默后的CNE2细胞增殖、克隆形成、迁移和侵袭能力均明显降低（P值均〈0.05）。结论 ：NONO基因沉默可抑制CNE2细胞的体外增殖、迁移及侵袭,提示NONO可能是一个潜在的促进鼻咽癌发生和发展的重要调控蛋白。

英文摘要：

Objective： To investigate the effects of non-POU domain containing octamer-binding （NONO） gene silencing on the proliferation, migration and invasion of nasopharyngeal carcinoma cells in vitro, in order to explore the role of NONO in the progression of nasopharyngeal carcinoma.Methods： The expression of NONO protein in normal nasopharyngeal epithelial cell line NP-69 and nasopharyngeal carcinoma cell lines CNE1 and CNE2 was detected by Western blotting. The lentiviruses containing NONO gene- specific shRNA were infected into CNE2 cells, then the stably infected CNE2 cell line was screened by puromycin. The efficiency of NONO gene silencing was determined by Western blotting. The proliferation, clone formation, migration and invasion abilities of NONO gene- silenced CNE2 cells were detected by Cell Counting Kit-8 （CCK-8）, plate clone formation experiment, wound healing assay as well as Transwell migration and invasion assay in vitro, respectively. Results： The expressions of NONO protein in nasopharyngeal carcinoma CNE1 and CNE2 cells were higher than that in normal nasopharyngeal epithelial NP-69 cells （both P 〈 0.05）. The silencing efficiency of NONO gene in CNE2 cells was close to 100% after infection with the lentiviruses containing NONO gene-specific shRNA. Compared with uninfected or negative control lentivirus-infected CNE2 cells, the proliferation, clone formation, migration and invasion of NONO gene-silenced CNE2 cells were significantly decreased （all P 〈 0.05）. Conclusion： NONO gene-silencing of can inhibit the proliferation, migration and invasion of CNE2 cells in vitro, which indicates that NONO may be a potential regulatory protein to promote the occurrence and development of nasopharyngeal carcinoma.