慢病毒介导RNA干扰HES1鼻咽癌稳定细胞株的建立与鉴定

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LV...

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Bibliographic Details
Published in山东医药 Vol. 57; no. 18; pp. 9 - 11
Main Author 程玉霜 王慧艳 肖东
Format Magazine Article
LanguageChinese
Published 南方医科大学肿瘤研究所,广州,510515 2017
Subjects
NPC细胞株
RNA干扰
Split多毛增强子1
慢病毒载体
鼻咽癌
RNA干扰
nasopharyngeal carcinoma
hairy and enhancer of split homolog-1
Split多毛增强子1
RNA interference
NPC cells
慢病毒载体
鼻咽癌
NPC细胞株
lentivirus expression vector
Online AccessGet full text
ISSN1002-266X
DOI10.3969/j.issn.1002-266X.2017.18.003

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Summary: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。
Bibliography:37-1156/R
nasopharyngeal carcinoma; hairy and enhancer of split homolog-1; RNA interference; lentivirus expression vector; NPC cells
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 gr
ISSN:1002-266X
DOI:10.3969/j.issn.1002-266X.2017.18.003