摘要
目的:探讨Survivin反义核酸对乳腺癌细胞增殖的影响.方法:本实验分6组,分别以脂质体介导反义寡核苷酸、无义寡核苷酸及RPMI 1640培养液作为空白的对照组,转染人乳腺癌MCF-7细胞系,采用Real time RT-PCR,Western blot方法检测各组MCF-7细胞Survivin蛋白及mRNA的表达情况,测定转染后细胞贴壁率变化,研究Survivin反义寡核苷酸对MCF-7细胞的生长抑制作用.结果:脂质体介导Sur-vivin反义寡核苷酸转染人乳腺癌MCF-7细胞后,细胞内Survivin蛋白及mRNA表达显著降低,通过Realtime RT-PCR检测MCF-7细胞经作用后其Survivin mRNA起始拷贝数明显下降;同时细胞贴壁率降低达32.96%,细胞生长抑制率最高达73.62%,ASOND/LiP组与NODN/LiP组和LiP组差异有显著性(P<0.05).结论:脂质体介导转染的反义寡核苷酸可在Survivin蛋白及mRNA水平有效地降低乳腺癌MCF-7细胞内Survivin的表达,并能够有效抑制乳腺癌细胞的增殖活性,提示Survivin靶向治疗可能成为乳腺癌综合治疗的一种重要方法.
Objective： To explore the influence of cell proliferation on human breast cancer after survivin was blocked with the antisense technique. Methods： The experiment was done in 6 groups. Antisense oligonucleotide （ASODN）, nonsense oligonucleotide （NODN） were transfected in human breast cancer MCF -7 cell line and RPMI 1640 culture medium （normal control）. Real time RT- PCR and western blot were applied in this study to detect and measure survivin protein and rnRNA expression in MCF- 7 cell. The cell adherent rate after transfection was measured to investigate the effect of antisense oligonucleotide on MCF- 7 cell growth restriction. Results： After survivin antisense oligonucleotide was transfected in human breast can- cer MCF- 7 cell, the expression of survivin protein and rnRNA decreased notably. Meanwhile, the cell adhcrcnt rate was reduced to 32.96 %, and the cell growth restriction rate could be as high as 73.62 %. There were statistical differences among the ASOND/Lip group and Lip group （P 〈 0.05）. Conclusion： ASODN can significantly reduce survivin expression in MCF - 7 cell of breast cancer at level of protein and mRNA and effectively suppress the proliferation of MCF - 7 cell when Lipofectin was used to transfect antisense oligonu- cleotide in survivin mRNA. Survivin target therapy may be used as an important method in breast cancer treatment.
出处
《昆明医学院学报》
2005年第3期45-50,共6页
Journal of Kunming Medical College
