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siRNA对肺癌细胞株NCI-H460 bcl-2基因表达的影响
引用本文:胡海燕,张洹.siRNA对肺癌细胞株NCI-H460 bcl-2基因表达的影响[J].暨南大学学报,2005,26(2):174-179.
作者姓名:胡海燕  张洹
作者单位:暨南大学医学院血液病研究所,广东,广州,510632
基金项目:广东省自然科学重点基金资助 (0 2 1195 )
摘    要:目的:研究siRNA (smallinterferenceRNA)对大细胞肺癌细胞株NCI -H4 6 0bcl- 2基因表达的影响。方法:利用Ambion公司提供的设计软件和试剂盒,设计合成以bcl - 2基因为靶标的siRNA ,通过脂质体将合成的siRNA转入NCI-H4 6 0细胞株,设置转染bcl- 2反义药物G3139和空白两对照组。用MTT法检测siRNA对细胞生长的作用;流式细胞仪检测细胞周期的改变和Bcl- 2蛋白表达;RT -PCR检测bcl- 2mRNA水平。结果:siRNA组与对照组细胞存活率均有显著性差异(P <0 0 5 ) ;siRNA组bcl- 2的mRNA明显低于对照组和反义组(P <0 0 5 ) ;siRNA组Bcl- 2蛋白阳性率明显低于对照组和反义组,siRNA组以及反义组细胞阻滞于S期。结论:体外转录合成的siRNA可抑制NCI-H4 6 0细胞bcl- 2基因的表达,抑制率可达5 0 %以上。

关 键 词:小干扰RNA  bcl-2基因  大细胞肺癌细胞株  NCI—H460
文章编号:1000-9965(2005)02-0174-06
修稿时间:2004年9月3日

Effect of siRNA on bcl-2 expression in human lung cancer cell line NCI-H460
HU Hai-yan,ZHANG Yuan.Effect of siRNA on bcl-2 expression in human lung cancer cell line NCI-H460[J].Journal of Jinan University(Natural Science & Medicine Edition),2005,26(2):174-179.
Authors:HU Hai-yan  ZHANG Yuan
Abstract:Aim: To evaluate the effectiveness of small interference RNA (siRNA) on bcl-2 gene expression. Methods: With Ambion's software and kit, we designed and synthesized siRNA targeted bcl-2, which were transfected into human lung cancer cell lines NCI-H460 with Lipofectamine. The non-transfected cells and cells treated with antisense drug (G-3139) were taken as controls. MTT was used to detect the inhibitive rate of cells growth. Flowcytometry method was used to detect the change of cycling of cell and protein level. The inhibitive effect of siRNA on mRNA level was detected by RT-PCR. Results: The survival rate of cells of siRNA groups were significantly lower than ones of control groups at 24,48 and 96 h? As compared with antisense group, there is only statistic difference at 24 and 48 h. PCR showed that the bcl-2 band of siRNA groups on electrophoresis were fade-out as compared with other groups. The results of flowcytometry showed the positive rate of Bcl-2 protein in siRNA groups was significantly lower than other groups and the cells were blocked at S stage. Conclusion: The level of bcl-2 gene expression of NCI-H460 cells would be declined by transfected with siRNA and inhibitive rate reached more than 50%.
Keywords:small interference RNA(siRNA)  bcl-2 gene  NCI-H460
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