RNAi-mediated gene silencing of vascular endothelial growth factor C suppresses growth and induces apoptosis in mouse breast cancer in vitro and in vivo.
RNAi-mediated gene silencing of vascular endothelial growth factor C suppresses growth and induces apoptosis in mouse breast cancer in vitro and in vivo.
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DOI：
10.3892/ol.2016.5158
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发表时间：
2016-11
期刊：
影响因子：
2.9
通讯作者：
Mu RH
中科院分区：
文献类型：
--
作者：
Liu YC;Ma WH;Ge YL;Xue ML;Zhang Z;Zhang JY;Hou L;Mu RH
血管内皮细胞生长因子（VEGF）-C通过允许淋巴结转移和淋巴管生成等作用促进肿瘤发生。RNA干扰（RNAi）是一种抑制靶基因表达的新技术，可以提高癌症治疗的有效性。本研究旨在探讨VEGF-C RNAi对小鼠乳腺癌细胞凋亡和增殖的影响。设计并合成了总共三对靶向小鼠VEGF-C的小干扰RNA（siRNA），然后通过脂质体方法转染到4 T1细胞中。采用逆转录聚合酶链反应（RT-PCR）、Western blot、3-（4，5-dimethylthiazol-2-yl）-2，5-diphenyltetrazolium bromide法、Hoechst 33258染色和流式细胞术检测4 T1细胞VEGF-C表达、caspase-3蛋白表达及细胞增殖和凋亡情况。还对患有乳腺癌的BALB/c小鼠进行了体内实验。测量肿瘤重量和体积，并通过TUNEL测定来评估肿瘤组织中的凋亡细胞的数目。采用免疫组化法和酶联免疫吸附法检测肿瘤组织中VEGF-C的表达。结果表明，三对siRNA，特别是siV 2，显著降低4 T1细胞中VEGF-C mRNA和蛋白水平。siV 2被认为是最有效的siRNA，因此被选择用于后续实验。此外，体外研究表明，VEGF-C RNAi显著降低细胞生长，诱导凋亡，并上调切割的caspase-3蛋白的表达。通过瘤内注射siV 2降低了乳腺癌体内模型中的肿瘤重量和体积。抗肿瘤疗效与VEGF-C表达降低和细胞凋亡诱导增加相关。因此，本研究表明，VEGF-C RNAi抑制小鼠乳腺癌的生长在体外和体内，它可能是一种新的靶向治疗乳腺癌。
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 cells in vitro and in 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 performed in vitro to analyze VEGF-C expression, cleaved caspase-3 protein expression and 4T1 cell proliferation and apoptosis. Experiments were also conducted in vivo on 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 vitro studies 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 cancer in vivo models 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 growth in vitro and in vivo and that it may be a novel targeted therapy for breast cancer.
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DOI：
10.4161/cc.8.14.9101
发表时间：
2009-07-15
期刊：
Cell cycle (Georgetown, Tex.)
影响因子：
--
作者：
Kurenova EV;Hunt DL;He D;Fu AD;Massoll NA;Golubovskaya VM;Garces CA;Cance WG
通讯作者：
Cance WG
影响因子：
11.5
作者：
Kodama, Michiyo;Kitadai, Yasuhiko;Chayama, Kazuaki
通讯作者：
Chayama, Kazuaki
影响因子：
3.8
作者：
Kinoshita, J;Kitamura, K;Sugimachi, K
通讯作者：
Sugimachi, K
DOI：
10.1007/978-1-4939-1538-5_1
发表时间：
2015-01-01
期刊：
RNA INTERFERENCE: CHALLENGES AND THERAPEUTIC OPPORTUNITIES
影响因子：
--
作者：
Sioud, Mouldy
通讯作者：
Sioud, Mouldy
慢病毒介导的靶向 VEGF-C 的小干扰 RNA 抑制乳腺癌中的肿瘤淋巴管生成和生长
DOI：
10.1002/ar.20893
发表时间：
2009-05-01
影响因子：
2
作者：
Guo, Baoliang;Zhang, Yafang;Zhang, Jianguo
通讯作者：
Zhang, Jianguo