中文摘要：

目的建立稳定干扰Split多毛增强子1（HES1）的鼻咽癌（NPC）细胞株,为进一步探讨HES1对NPC的影响奠定基础。方法将293T细胞接种6孔板后随机分为A、B、C、D、E组,分别瞬时转染慢病毒质粒shHES1-a、shHES1-b、shHES1-c、shHES1-d及空载质粒shSCR,分别用qRT-PCR和Western blot法检测HES1 mRNA及蛋白,筛选出最有效干扰质粒;将最有效干扰质粒或空载质粒与慢病毒包装质粒共转染293T细胞,获得含最有效干扰质粒或空载质粒的病毒上清（LV-shHES1/LV-shSCR）。将NPC细胞CNE2和S18随机分为观察组和对照组,分别将LVshHES1、LV-shSCR以浓度梯度方式感染细胞,分别采用qRT-PCR和Western blot法检测HES1 mRNA及蛋白。结果C组293T细胞中HES1 mRNA及蛋白相对表达量均低于其余各组（P均〈0.05）。与对照组比较,观察组CNE2和S18细胞中HES1 mRNA及蛋白相对表达量均降低（P均〈0.05）。结论成功建立稳定干扰HES1的NPC细胞株。

英文摘要：

Objective To establish a nasopharyngeal carcinoma（ NPC） cell line for silencing hairy and enhancer of split homolog-1（ HES1）,and it will lay a foundation for further study on the effect of HES1 on NPC. Methods cell 293 T inoculated on a 6-well plate were divided into groups A,B,C,D and E,which were instantaneously transfected with the recombinant lentiviruses（ shHES1-a,shHES1-b,shHES1-c,shHES1-d and empty vector shSCR）. The expression of HES1 was analyzed by qRT-PCR and Western blotting to identify the LV-shHES1 vector that had the highest gene knockdown efficiency. cell 293 T were co-transfected with the most effective interference plasmid or no-load plasmid and lentiviral packaging plasmid to obtain the virus supernatant（ LV-shHes1/LV-shSCR） containing the most effective interference plasmid or no-load plasmid. NPC cells CNE2 and S18 were randomly divided into the observation group and control group. LVshHes1 and LV-shSCR were used to infect the cells in a concentration gradient manner,and the mRNA and protein of HES1 was detected by qRT-PCR and Western blot. Results The relative expression of HES1 mRNA and protein in 293 T cells in the group C was lower than that in the other groups,and the difference was statistically significant（ all P〈0. 05）.Therefore,the shHES1-c interference efficiency was the best. Compared with the control group,the relative expression of HES1 mRNA and protein in both CNE2 and S18 cells of the observation group was decreased（ all P〈0. 05）. Conclusion NPC cell line for silencing HES1 gene by RNA interference was successfully established.