title: RNAi-mediated gene silencing of vascular endothelial growth factor C suppresses growth and induces apoptosis in mouse breast cancerin vitroandin vivo.
abstract: Vascular endothelial cell growth factor (VEGF)-C promotes tumorigenesis by allowing lymph node metastasis and lymphangiogenesis, among other actions. RNA interference (RNAi) is a novel technique for suppressing target gene expression and may increase the effectiveness of cancer treatments. The present study assessed the influence of VEGF-C RNAi on the apoptosis and proliferation of mouse breast cancer cellsin vitroandin vivo. A total of three pairs of small interfering RNA (siRNA) targeting mouse VEGF-C were designed and synthesized prior to transfection into 4T1 cells via a liposomal approach. Reverse transcription polymerase chain reaction, western blot analysis, a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, Hoechst 33258 staining and flow cytometry were performedin vitroto analyze VEGF-C expression, cleaved caspase-3 protein expression and 4T1 cell proliferation and apoptosis. Experiments were also conductedin vivoon BALB/c mice with breast cancer. Tumor weight and volume were measured and the number of apoptotic cells in tumor tissues was assessed by a TUNEL assay. Immunohistochemical assays and an enzyme-linked immunosorbent assay were used to measure the expression of VEGF-C in tumor tissues. The results demonstrated that the three pairs of siRNA, particularly siV2, significantly reduced VEGF-C mRNA and protein levels in 4T1 cells. siV2 was deemed to be the most efficient siRNA and therefore was selected to be used in subsequent experiments. Furthermore,in vitrostudies indicated that VEGF-C RNAi significantly decreased cell growth, induced apoptosis and upregulated the expression of cleaved caspase-3 protein. Tumor weight and volume in breast cancerin vivomodels was reduced by the intratumoral injection of siV2. Antitumor efficacy was associated with decreased VEGF-C expression and increased induction of apoptosis. The present study therefore indicated that VEGF-C RNAi inhibited mouse breast cancer growthin vitroandin vivoand that it may be a novel targeted therapy for breast cancer.
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, Qingdao, Shandong 266021, P.R. China; Department of Immunology, Medical College, Beihua University, Jilin, Jilin 132013, P.R. China.
authors.forename: Yong-Chao
authors.initials: YC
authors.lastname: Liu
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, Qingdao, Shandong 266021, P.R. China.
authors.forename: Wen-Hui
authors.initials: WH
authors.lastname: Ma
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, Qingdao, Shandong 266021, P.R. China.
authors.forename: Yin-Lin
authors.initials: YL
authors.lastname: Ge
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, Qingdao, Shandong 266021, P.R. China.
authors.forename: Mei-Lan
authors.initials: ML
authors.lastname: Xue
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, Qingdao, Shandong 266021, P.R. China.
authors.forename: Zheng
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, Qingdao, Shandong 266021, P.R. China.
authors.forename: Jin-Yu
authors.initials: JY
authors.lastname: Zhang
authors.affiliation: Department of Biochemistry and Molecular Biology, Medical College, Qingdao University, Qingdao, Shandong 266021, P.R. China.
authors.forename: Lin
authors.initials: L
authors.lastname: Hou
authors.affiliation: Department of Immunology, Medical College, Beihua University, Jilin, Jilin 132013, P.R. China.
authors.forename: Run-Hong
authors.initials: RH
authors.lastname: Mu
keywords: RNA interference; apoptosis; breast cancer; small interfering RNA; vascular endothelial growth factor-C