Proliferation suppression and apoptosis of ovarian carcinoma cells induced by small interfering RNA against vascular endothelial growth factor.
OBJECTIVE
The aim of this study was to investigate whether RNA interference (RNAi) targeting vascular endothelial growth factor (VEGF) could inhibit the proliferation and induce apoptosis of ovarian carcinoma cells.


METHODS
Human epithelial ovarian carcinoma cell line CaoV3 was transfected with VEGF-targeted small interfering RNA (siRNA) for 48 h. The down-regulation of VEGF expression was determined by real-time polymerase chain reaction and Western blot. The proliferation of CaoV3 cells was detected by a colorimetric BrdU assay. Caspase-3 activity and TUNEL assay were also detected to study the apoptosis of ovarian carcinoma cells induced by VEGF siRNA. The protein levels of survivin, MMP2 and MMP9 were measured by Western blot.


RESULTS
mRNA and protein of VEGF were significantly down-regulated by VEGF siRNA. Down-regulation of VEGF expression dramatically suppressed the proliferation of CaoV3 cells, and increased the Caspase-3 activity. Nearly 100% of cells indicated TUNEL-positive. The expression of anti-apoptotic protein survivin was decreased, and the invasive related protein MMP2 and MMP9 were also significantly reduced by VEGF siRNA.


CONCLUSIONS
Our study implied that siRNA targeting VEGF could effectively inhibit cell proliferation, induce cell apoptosis, and decrease the cell invasive potential. These findings suggest that the RNAi approach targeting VEGF may be an effective therapeutic strategy for ovarian cancer.
Zhou
Jianwei
J
Department of Gynecology, The Second Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang Province, China.
Gan
Ningyue
N
Zhang
Weijiang
W
Lu
Weiguo
W
Xie
Xing
X
D000704:Analysis of Variance; D017209:Apoptosis; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002478:Cells, Cultured; D015536:Down-Regulation; D005260:Female; D006801:Humans; D020287:In Situ Nick-End Labeling; D051034:Inhibitor of Apoptosis Proteins; D020778:Matrix Metalloproteinase 2 / Q000235:genetics / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000235:genetics / Q000378:metabolism; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D010053:Ovary / Q000166:cytology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077022:Survivin; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
C107489:BIRC5 protein, human; D051034:Inhibitor of Apoptosis Proteins; D008869:Microtubule-Associated Proteins; D034741:RNA, Small Interfering; D000077022:Survivin; D042461:Vascular Endothelial Growth Factor A; C522361:MMP2 protein, human; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9