山东医药2017，Vol.57Issue(18)：9-11,3.DOI:10.3969/j.issn.1002-266X.2017.18.003
慢病毒介导RNA干扰HES1鼻咽癌稳定细胞株的建立与鉴定
Establishment and identification of NPC cell line for silencing HES1 gene by RNA interference
摘要
Abstract
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 293T 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 293T 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.LV-shHes1 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 293T 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.关键词
鼻咽癌/Split多毛增强子1/RNA干扰/慢病毒载体/NPC细胞株Key words
nasopharyngeal carcinoma/hairy and enhancer of split homolog-1/RNA interference/lentivirus expression vector/NPC cells分类
医药卫生引用本文复制引用
程玉霜,王慧艳,肖东..慢病毒介导RNA干扰HES1鼻咽癌稳定细胞株的建立与鉴定[J].山东医药,2017,57(18):9-11,3.基金项目
国家自然科学基金资助项目(81172587,81372896,81672689) （81172587,81372896,81672689）
广东省自然科学基金资助项目(2014A030313294). （2014A030313294）
