Effects of RNAi-silenced livin gene on glioma cell lines U251

LAN Hu-xiang; et al

doi:10.13406/j.cnki.cyxb.2010.08.029

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Effects of RNAi-silenced livin gene on glioma cell lines U251

更新时间：2024-01-17

- Effects of RNAi-silenced livin gene on glioma cell lines U251
- Journal of Chongqing Medical University Vol. 35, Issue 8, Pages: 1145-1148(2010)
- 作者机构：
  
  重庆医科大学附属第一医院神经外科
- 作者简介：
- 基金信息：
- DOI：10.13406/j.cnki.cyxb.2010.08.029
  CLC： R739.4
- Published：2010，
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[1]兰胡翔,霍钢,郑履平.RNAi沉默livin基因表达对胶质瘤U251细胞增殖的影响[J].重庆医科大学学报,2010,35(08):1145-1148.

LAN Hu-xiang, et al. Effects of RNAi-silenced livin gene on glioma cell lines U251. [J]. Journal of Chongqing Medical University 35(8):1145-1148(2010)
[1]兰胡翔,霍钢,郑履平.RNAi沉默livin基因表达对胶质瘤U251细胞增殖的影响[J].重庆医科大学学报,2010,35(08):1145-1148. DOI： 10.13406/j.cnki.cyxb.2010.08.029.

LAN Hu-xiang, et al. Effects of RNAi-silenced livin gene on glioma cell lines U251. [J]. Journal of Chongqing Medical University 35(8):1145-1148(2010) DOI： 10.13406/j.cnki.cyxb.2010.08.029.

摘要

目的:通过RNA干扰（RNA interference

RNAi）阻断胶质瘤U251细胞中livin基因的表达

研究livin基因沉默后对细胞生长和细胞凋亡的影响。方法:将合成的DNA正义链及反义链变性、退火

形成的双链DNA与pSilencer2.0线性质粒以T4DNA连接酶连接

重组质粒经酶切鉴定后

用脂质体法转染胶质瘤U251细胞

通过RT-PCR、免疫印迹检测干扰效果。MTT法检测细胞生长的影响

流式细胞仪检测转染后胶质瘤U251细胞的凋亡变化。结果:酶切证实设计合成的DNA片段已正确插入载体。RT-PCR、免疫印迹检测实验表明:pSilencer2.0-liv1、pSilencer2.0-liv2重组载体均能有效地阻断胶质瘤U251细胞中livin基因的表达

使livin基因在mRNA和蛋白水平上的表达明显下调（P<0.01）

后者抑制效果更佳。转染后的U251细胞生长速度明显变慢。转染pSilencer2.0-liv2实验组细胞的凋亡增加了17.0%。结论:重组真核载体能有效抑制U251细胞的livin表达

抑制细胞的增殖

诱导细胞凋亡。

Abstract

Objective:To construct eukaryotic expression vectors by usingthep Silencer2.0 vector for inhibiting human livin gene by RNA interference

and to detect the effect of the silenced livin gene on the growth and apoptosis of U251 cells. Methods:Two target gene segments were synthesized and cloned into the pSilencer2.0 vectorrespectively to construct two recombinant eukaryotic expression vectors: pSilencer2.0-liv1 and pSilencer2.0-liv2

both of which were identified by enzyme digestion analysis. Then the U251 cells were transfected with the recombinant vectors and the interfering effects were detected by RT-PCR and Western blot. The proliferation of the U251 cells was detected by MTT method and their apoptosis index detected by flow cytometry. Results:The results of Enzyme digestion analysis showed that two target segments were cloned into pSilencer2.0 vectors. The results of RT-PCR and Western blot indicated that pSilencer2. 0-liv1 and pSilencer2.0-liv2 vectors could knock down the transcription and expression of livin gene

but the effect of pSilencer2.0-liv2 vectors was better than that of the pSilencer2.0-liv1 vectors. Obvious decrease in growth of the U251 cells was observed after transfected with pSilencer2.0-liv1 and pSilencer2.0-liv2 vectors

along with 17%increase of the apoptosis index in the U251 cells transfected with pSilencer2.0-liv2 vectors. Conclusion:The transcription and expression of livin gene were inhibited effectively in the glioma cell line U251. Aftertransfected with pSilencer2.0-liv2 vectors

the apoptosis index of the U251 cells was increased and the growth inhibited.

关键词

RNA干扰livin胶质瘤细胞

Keywords

RNA interferencelivinglioma cell

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