RNAi沉默肺癌细胞中VEGF的表达治疗肺癌机制的初步研究
Mechanism of RNAi silence inhibiting VEGF on therapy of lung cancer
张泳 1谷仲平 2周勇安 2王云杰2
作者信息
- 1. 第四军医大学唐都医院胸外科,陕西,西安,710038;陕西中医学院附属医院胸外科,陕西,咸阳,712000
- 2. 第四军医大学唐都医院胸外科,陕西,西安,710038
- 折叠
摘要
目的:探讨RNAi干涉沉默肺癌细胞中VEGF的表达治疗肺癌机制.方法:将psilencerTM-3.1-VEGF分别稳定转染人人肺癌细胞株A549,建立稳转细胞系.通过RT-PCR检测Bel-2和Bax的mRNA的水平.免疫印迹方法检测稳转的人肺癌细胞株A549中P53,Bcl-2和Bax的表达.流式细胞仪检测稳转的人肺癌细胞株A549细胞系的凋亡.结果:稳定转染pSilenceTM-3.1-VEGF的人肺癌细胞株A549细胞系中VEGF的表达明显下调.稳转的人肺癌细胞株A549细胞系中P53,Bcl-2和Bax的表达无明显变化,未观察到明显的细胞凋亡现象.与对照组比较,在人HUVEC细胞系的作用下,稳定转染的人A549细胞虽Bax的表达未发生变化,但P53表达明显增高,Bcl-2表达下调,并且呈现细胞凋亡现象.外源性VEGF能够增加Bcl-2的表达,并导致P53表达下调.结论:RNAi沉默肺癌细胞中VEGF的表达未影响肺癌细胞自身的变化,其对肺癌发展的抑制可能是通过抑制了血管内皮细胞上的VEGF/VEGFR通路,从而引起了肺癌细胞的凋亡.
Abstract
AIM: To research the mechanism of therapy of cancer with RNAi inhibiting VEGF. METHODS: The pSilencerTM-3.1-VEGF vector was thansfected into the A549 cell line. The mRNA level of Bcl-2 and Bax were detected by RT-PCR. And the expreesion of P53, Bcl-2 and Bax in stable thansfected A549 were analysized by western blot. The apoptosis of stable thasfected A549 was estimated with FCM. RESULTS: The expression of VEGF in the stable thansfected A549 was inhibited obviously. Meanwhile, the P53 , Bcl-2 and Bax expression did not change in the stable transfected AS49 cell lines. And we did not observe the apoptosis of stable transfected A549 cells. Compared with the control group, althrough the Bax expression did not change in the stable transfected A549 cell lines under the HUVEC, the P53 expression increased, while the Bcl-2 decreased, at the sametime, we characterized the apoptosis of transfected A549. What's more, this phenomina could be reversed with the VEGF. CONCLUSION: The mechanism of therapy of cancer with RNAi inhibiting VEGF could result from the blockade the VEGF/VEGFR.
关键词
RNAi/VEGF/肺癌/机制Key words
RNAi/VEGF/lung carcinoma/mechanism引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据