RNA干涉抑制存活素表达并诱导HeLa S3细胞凋亡的研究

RNA干涉抑制存活素表达并诱导HeLa S3细胞凋亡的研究

王瑞瑛¹,梅柱中²,董燕²,钱俊杰²,赵国伟¹,田宝磊²,董申¹,罗贵民¹,孙志贤²

（1.吉林大学分子酶学工程教育部重点实验室，长春130021；2.军事医学科学院放射医学研究所，北京100850）

收稿日期:2005-03-18 修回日期:1900-01-01 出版日期:2005-09-26 发布日期:2005-09-26
通讯作者: 孙志贤

Inhibition of Survivin Expression and Induction of Apoptosis of HeLa S3 Cells by RNA Interference

WANG Rui-ying¹,MEI Zhu-zhong²,DONG Yan²,QIAN Jun-jie²,ZHAO Guo-wei¹,TIAN Bao-lei²,DONG Shen¹,LUO Gu-min¹,SUN Zhi-xian²

（1．Key Laboratory for Molecular Enzymology and Engineering of the Ministry of Education，Jilin University，Changchun130021，China；2.Institute of Radiation Medicine，Academy of Military Medical Sciences，Beijing 100850，China）

Received:2005-03-18 Revised:1900-01-01 Online:2005-09-26 Published:2005-09-26
Contact: SUN Zhi-xian

摘要/Abstract

摘要： 选择3个靶向存活素（S1，S2，S3）Survivin基因的siRNA序列，构建相应的RNA干涉载体pTet-U6-S1，pTet-U6-S2，pTet-U6-S3，将它们分别转染HeLa S3细胞，采用RT-PCR和Western印迹检测分析转染对HeLa S3细胞内源Survivin表达的影响，结果表明，针对Survivin 3’端非编码区序列的干涉载体pTet-U6-S3转染细胞后，Survivin的mRNA水平和蛋白水平均明显下调，抑制水平高于S1序列.与对照相比，S1，S2和S3片段对Survivin mRNA的抑制率分别是20%，12.5%和40%，对Survivin蛋白的抑制率分别是39.2%，17.0%和58.6%. 流式细胞术检测结果表明，抑制Survivin表达后，HeLa S3细胞凋亡比例显著提高.

关键词: 存活素, RNA干涉, 凋亡, HeLaS3细胞

Abstract: To study the functions of survivin gene further，three siRNA segments （S1，S2,，S3）specifically targeting survivin were selected.Accordingly RNAi plasmids pTet-U6-S1，pTet-U6-S2，pTet-U6-S3 were constructed.The changes of survivin expression were examined by semi-quantitative reverse transcription polymerase chain reaction （RT-PCR）and Western-blot analysis after the plasmids had been transfected into HeLa S3.The results show that survivin expression is inhibited at both mRNA and protein levels after the plasmid pTet-U6-S3 has been transfected,，which targets on the 3’non-encoding region of surviving.The inhibition level is much higher than that of S1 sequence reported previously.Compared with that of the control，the inhibition ratios of S1，S2，S3 to mRNA of survivin are 20%，12.5% and 40% respectively，and 39.2%，17.0% and 58.6% to its protein.And the effect of the plasmid pTet-U6-S3 on the cell apoptosis was analyzed by flow cytometry.The apoptosis of HeLa S3 cells was greatly increased after transfection for 48h.

Key words: survivin, RNA interference, apoptosis, HeLa S3 cell

中图分类号:

王瑞瑛,梅柱中,董燕,钱俊杰,赵国伟,田宝磊,董申,罗贵民,孙志贤. RNA干涉抑制存活素表达并诱导HeLa S3细胞凋亡的研究[J]. J4, 2005, 43(05): 691-695.

WANG Rui-ying,MEI Zhu-zhong,DONG Yan,QIAN Jun-jie,ZHAO Guo-wei,TIAN Bao-lei,DONG Shen,LUO Gu-min,SUN Zhi-xian. Inhibition of Survivin Expression and Induction of Apoptosis of HeLa S3 Cells by RNA Interference[J]. J4, 2005, 43(05): 691-695.

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