Effects of RNAi targeting Livin on proliferation and apoptosis of MDA-MB-231 cells

LI Fan; WANG Xiao-yi; CHEN Li; TIAN Tian; LI Jia; REN Guo-sheng

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Effects of RNAi targeting Livin on proliferation and apoptosis of MDA-MB-231 cells

更新时间：2025-01-15

- Effects of RNAi targeting Livin on proliferation and apoptosis of MDA-MB-231 cells
- Journal of Army Medical University Vol. 30, Issue 24, Pages: 2302-2306(2008)
- 作者机构：
  
  重庆医科大学附属第一医院普外科
- 作者简介：
- 基金信息：
- DOI：
  CLC： R329.25;R394.3;R737.9
- Published：2008，
  
  Published Online：30 December 2008，
- 稿件说明：
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LI FAN, WANG XIAO-YI, CHEN LI, et al. Effects of RNAi targeting Livin on proliferation and apoptosis of MDA-MB-231 cells. [J]. Journal of army medical university, 2008, 30(24): 2302-2306.
DOI：

LI FAN, WANG XIAO-YI, CHEN LI, et al. Effects of RNAi targeting Livin on proliferation and apoptosis of MDA-MB-231 cells. [J]. Journal of army medical university, 2008, 30(24): 2302-2306. DOI：

摘要

目的研究凋亡抑制蛋白Livin靶向RNA干扰MDA-MB-231对细胞增殖和凋亡的影响，探讨其可能的机制。方法构建3个靶向Livin的siRNA真核表达载体，分别转染乳腺癌细胞MDA-MB-231后，通过RT-PCR、Western blot技术检测干扰前后MDA-MB-231细胞Livin、Caspase-3和Smac/DIABLO在mRNA、蛋白水平的表达；采用MTT法检测对细胞增殖的抑制作用，流式细胞法测细胞周期，TUNEL法检测细胞凋亡以及观察细胞形态变化。结果成功构建了针对Livin基因的3个特异性siRNA表达载体。它们不同程度地抑制了MDA-MB-231细胞Livin基因的表达，以si-Livin2抑制效率最高，它对Livin的mRNA和蛋白的抑制率分别达76.4%，70.2%。si-Livin2转染48 h后，与未转染组相比，细胞增殖活力受到抑制，转染48 h抑制率达36.1％。细胞周期重新分布，G

期上升为(71.75±1.31)％，S期下降为(18.06±1.46)％；细胞凋亡率上升为(13.28±1.65)％；电镜下可见典型的凋亡细胞特征。细胞内Caspase-3

Smac/DIABLO的mRNA、蛋白表达有所上升。结论 Livin靶向RNA干扰通过上调Caspase-3

Smac/DIABLO表达有效地抑制MDA-MB-231细胞增殖，促进凋亡。

Abstract

Objective To study the mechanisms by which the RNA targeting Livin regulates the proliferation and apoptosis of MDA-MB-231 cells. Methods Three siRNA eukaryotic expression vectors targeting Livin were constructed and transfected into breast cancer MDA-MB-231 cells. The changes of Livin

Caspase-3 and Smac/DIABLO expressions in MDA-MB-231 cells with or without RNAi transfection were detected by quantitative

Real-Time PCR and Western blotting. The proliferation of MDA-MB-231 cells was detected by MTT assay

their cell cycles by flow cytometry and their apoptosis by TUNEL assay and their morphological changes observed by optical microscope and electron microscope. Results Three specific siRNA eukaryotic expression vectors targeting Livin were successfully constructed. They all could inhibit the expressions of Livin mRNA and protein to different degree

of which si-Livin2 showed the highest inhibition rate. Forty-eight hours after si-Livin2 transfection

the proliferation of MDA-MB-231 cells was strongly inhibited

with the inhibition rate of 36.1%. The cell percentage of the G

phase was increased and that of the S phase was decreased. The apoptosis rate of MDA-MB-231 cells increased and the typical apoptosis was observed under electron microscope. The mRNA and protein expressions of Caspase-3 and Smac/DIABL were all increased. Conclusion Through upregulating the expressions of Caspase-3 and Smac/DIABLO

RNAi targeting Livin inhibits the proliferation and induces the apoptosis of MDA-MB-231 cells.

关键词

乳腺癌LivinRNA干扰增殖凋亡MDA-MB-231细胞株

Keywords

breast cancerLivinRNA interferenceproliferationapoptosisMDA-MB-231 cell line

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