慢病毒介导的Nestin基因沉默抑制人食管癌ECA109细胞增殖

基础医学与临床 ›› 2015, Vol. 35 ›› Issue (9): 1214-1218.

慢病毒介导的Nestin基因沉默抑制人食管癌ECA109细胞增殖

曹燕妮¹,王华川¹,王聪懿²,朱深银¹,温剑虎¹

1. 重庆医科大学附属第一医院
2. 重庆医科大学

收稿日期:2015-02-11 修回日期:2015-04-24 出版日期:2015-09-05 发布日期:2015-09-07
通讯作者: 温剑虎 E-mail:tiger_w02@163.com
基金资助:
重庆市卫生局重点科研项目

Silencing Nestin expression mediated by lentivirus inhibits human esophageal cancer ECA109 cell proliferation

Received:2015-02-11 Revised:2015-04-24 Online:2015-09-05 Published:2015-09-07

摘要/Abstract

摘要： 目的通过体外实验探讨沉默巢蛋白（Nestin）基因对人食管癌ECA109细胞增殖能力的影响及可能机制。方法合成Lenti-Nestin慢病毒载体，实验设blank组、scrambled组和Lent-Nestin组，转染ECA109细胞，建立稳定沉默Nestin基因的细胞株Lenti-Nestin；用qRT-PCR和Western blot检测Nestin、c-myc和cyclinD1 mRNA和蛋白表达；CCK-8法检测细胞增殖能力；平板集落实验检测细胞的集落形成。结果稳定沉默Nestin的细胞株构建成功；与blank组和scrambled组相比，Lent-Nestin组中Nestin mRNA（P<0.01）和蛋白（P<0.05）水平明显降低；细胞增殖能力明显降低（P<0.01），集落形成能力显著下降（P<0.05）；沉默Nestin表达后，c-myc、cyclinD1表达明显受到抑制(P<0.05)。结论沉默Nestin基因表达可抑制人食管癌ECA109细胞的增殖能力，其可能通过调控c-myc、cyclinD1表达参与其中。

关键词: Nestin, 食管癌, 增殖, 基因沉默

Abstract: Objective To investigate the effect of Nestin gene silencing on the proliferation of human esophageal cancer ECA109 cells and possible mechanism. Methods Lenti-Nestin was constructed and transfected into ECA109 cells to establish a stable Nestin-silencing cell line Lenti-Nestin. Blank group, scrambled group, Lenti-Nestin group were set up. The expressions of Nestin, c-myc and cyclinD1 mRNA and protein levels were detected by qRT-PCR and Western blot. The cell proliferation was analyzed by CCK8 assay. Results The stable Nestin-silencing cell line was successfully established. The expression of Nestin mRNA (P<0.01) and protein (P<0.05) levels were reduced significantly and the downregulation evidently suppressed cell proliferation (P<0.01) and colony forming capacity (P<0.05) compare with the scrambled group and blank group. However，The c-myc and cyclinD1 expression levels in ECA109 cells in Lenti-Nestin group were significantly lower than that of scrambled group and blank group (P<0.05). Conclusion Knockdown Nestin expression significantly inhibits the level of esophageal cancer ECA109 cells proliferation，which may act via influencing the expression of c-myc, cyclinD1．

Key words: Nestin, esophageal, proliferation, gene silencing

中图分类号:

R735.1

曹燕妮王华川王聪懿朱深银温剑虎. 慢病毒介导的Nestin基因沉默抑制人食管癌ECA109细胞增殖[J]. 基础医学与临床, 2015, 35(9): 1214-1218.

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