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


CONCLUSIONS
The siRNA targeting human VEGF gene could reduce VEGF expression, inhibit cellular proliferation and induce apoptosis in MCF-7 cells in vitro.
Zhang
Xiao
X
Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, Qingdao 266021, China.
Xu
Wen-hua
WH
Ge
Yin-lin
YL
Hou
Lin
L
Li
Quan
Q
D017209:Apoptosis / Q000235:genetics; D001943:Breast Neoplasms; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006801:Humans; D007150:Immunohistochemistry; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
D034741:RNA, Small Interfering; D042461:Vascular Endothelial Growth Factor A