陆志斌,周存荣,郑琳,张爱平,徐晓晗,吴卫兵,曲晨
LU Zhibin,ZHOU Cunrong,ZHENG Lin,ZHANG Aiping,XU Xiaohan,WU Weibing,QU Chen

• 1:浦口区中心医院心胸外科
• 2:南京市第一医院心胸外科
• 3:江苏省人民医院心胸外科
• 4:南京医科大学第二附属医院

摘要(Abstract)：

目的:构建DKK1的靶向小干扰RNA(siDKK1),探讨siRNA干扰DKK1表达后对食管癌ECA109细胞ALDH1A1 mRNA及蛋白水平的影响及机制。方法:ECA109细胞分为阴性对照组和siDKK1组,siDKK1组瞬时转染DKK1siRNA,RT-PCR及Western blot分别检测转染前后DKK1和ALDH1A1的mRNA及蛋白水平,Western blot检测JNK及c-Jun的磷酸化修饰水平。结果:与阴性对照组相比,siDKK1组ECA109细胞中DKK1和ALDH1A1的mRNA和蛋白表达水平都受到明显抑制(P<0.05),而调控ALDH1A1表达的上游因子JNK及c-Jun的磷酸化修饰水平都明显降低(P<0.05)。结论:DKK1 siRNA可能通过抑制JNK及c-Jun的磷酸化修饰从而下调ALDH1A1 mRNA和蛋白的表达。
Objective: To construct DKK1-targeted small interfering RNA( siDKK1) and to investigate the effect and mechanism of siDKK1 interference on the expression of ALDH1A1 mRNA and protein in esophageal carcinoma cell line ECA109. Methods: ECA109 cells were divided into negative control group and siDKK1 group. DKK1 siRNA was transiently transfected into siDKK1 group cells. RTPCR and Western blot were adopted to detect DKK1 and ALDH1A1 mRNA expression and protein expression levels before and after transfection. Western blot was adopted to detect phosphorylation level of JNK and c-Jun. Results: Compared with negative control group,the mRNA and protein expression level of DKK1 and ALDH1A1 in ECA109 cells of siDKK1 group was significantly inhibited( P < 0. 05),while phosphorylation level of upstream factor JNK and c-Jun that modulate the expression of ALDH1A1 decreased significantly( P < 0. 05). Conclusion: DKK1 siRNA may effectively inhibit the mRNA and protein expression of DKK1 by inhibiting the phosphorylation of JNK and c-Jun.

关键词(KeyWords)： 食管肿瘤;RNA干扰;基因表达;蛋白抑制
esophageal neoplasms;RNA interference;gene expression;protein inhibition

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金青年项目(81501202);; 南京市医学科技发展项目(YKK14182);; 南京医科大学科技发展基金项目(2014NJMU167)

作者(Author): 陆志斌,周存荣,郑琳,张爱平,徐晓晗,吴卫兵,曲晨
LU Zhibin,ZHOU Cunrong,ZHENG Lin,ZHANG Aiping,XU Xiaohan,WU Weibing,QU Chen

DOI: 10.19367/j.cnki.1000-2707.2017.04.015

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