[靶向VEGF基因的siRNA转染对人乳腺癌细胞增殖和凋亡的影响]

[Effect of siRNA transfection targeting VEGF gene on proliferation and apoptosis of human breast cancer cells].

作者信息

Zhang Xiao, Xu Wen-hua, Ge Yin-lin, Hou Lin, Li Quan

机构信息

Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, Qingdao 266021, China.

出版信息

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2007 Jan;23(1):14-7.

PMID:17210097

Abstract

AIM

To explore the effect of RNA interference-mediated vascular endothelial growth factor (VEGF) gene silencing on proliferation and apoptosis of human breast cancer MCF-7 cells.

METHODS

Small interfering RNA (siRNA) targeting VEGF gene was designed and DNA template was synthesized, then siRNA was obtained by in vitro transcription. After ds-siRNA was transfected into MCF-7 cells with Lipofectamine, the proliferation of MCF-7 cells was detected by MTT colorimetry, and the apoptosis was measured by Hoechst33258 staining. The change of cell cycle was analyzed by flow cytometry (FCM). The expression of VEGF on mRNA and protein level was analyzed by RT-PCR and immunohistochemical technology respectively.

RESULTS

The siRNA targeting human VEGF gene effectively inhibited the proliferation of MCF-7 cells, induced cell apoptosis, arrested the cell cycle in G(0)/G(1) phase, and decreased the expression of VEGF on both mRNA and protein level. But these effects did not appear in scrambled siRNA (siRNA(SCR)) control group.

CONCLUSION

The siRNA targeting human VEGF gene could reduce VEGF expression, inhibit cellular proliferation and induce apoptosis in MCF-7 cells in vitro.

摘要

目的

探讨RNA干扰介导的血管内皮生长因子（VEGF）基因沉默对人乳腺癌MCF-7细胞增殖和凋亡的影响。

方法

设计针对VEGF基因的小干扰RNA（siRNA）并合成DNA模板，然后通过体外转录获得siRNA。用脂质体将双链siRNA转染至MCF-7细胞后，采用MTT比色法检测MCF-7细胞的增殖情况，用Hoechst33258染色法检测细胞凋亡情况。通过流式细胞术（FCM）分析细胞周期的变化。分别采用逆转录-聚合酶链反应（RT-PCR）和免疫组织化学技术分析VEGF在mRNA和蛋白水平的表达。