Lentivirus-Mediated Small Interfering RNA Targeting VEGF-C Inhibited Tumor Lymphangiogenesis and Growth in Breast Carcinoma
Lentivirus-Mediated Small Interfering RNA Targeting VEGF-C Inhibited Tumor Lymphangiogenesis and Growth in Breast Carcinoma
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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
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发表时间：
2009-05-01
影响因子：
2
通讯作者：
Zhang, Jianguo
中科院分区：
文献类型：
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作者：
Guo, Baoliang;Zhang, Yafang;Zhang, Jianguo
淋巴结转移是影响乳腺癌患者预后的主要因素。血管内皮生长因子-C的激活通过促进淋巴管生成在淋巴结转移中起关键作用。因此，我们试图阐明针对血管内皮生长因子-C的小干扰RNA(SiRNA)是否能够抑制体内淋巴管的生成和淋巴结转移。以慢病毒为基础的血管内皮生长因子C siRNA载体感染乳腺癌细胞和异种移植瘤模型。采用实时定量聚合酶链式反应(QRT-PCR)、免疫组织化学和免疫印迹等方法检测血管内皮细胞生长因子C(VEGF-C)mRNA和蛋白的表达。采用侵袭实验检测血管内皮细胞生长因子C siRNA对乳腺癌细胞的影响。免疫组织化学方法检测淋巴管生成情况。慢病毒介导的血管内皮细胞生长因子C siRNA稳定地降低了血管内皮细胞生长因子C的mRNA和蛋白表达。血管内皮生长因子C siRNA抑制乳腺癌细胞的侵袭能力。体外培养。瘤内注射5周后，在MDA-MB-231细胞移植模型中，VEGF-C siRNA组的肿瘤体积明显小于对照scurble siRNA组。VEGF-C siRNA组LYVE-1和D2-40阳性血管数明显减少，提示VEGF-C siRNA抑制淋巴管生成。此外，体内应用血管内皮生长因子C siRNA可显著抑制肿瘤的淋巴结转移。综上所述，这些结果表明，慢病毒介导的VEGF-C siRNA为预防乳腺癌生长和淋巴转移的治疗干预提供了一种新的途径。Anat Rec，292：633-639,2009。(C)2009年Wiley-Liss，Inc.
Lymph node metastasis is a major prognostic factor for Patients with breast cancer. The activation of vascular endothelial growth factor (VEGF)-C plays a key role in lymph node metastasis through promoting lymphangiogenesis. Thus, we attempted to elucidate whether small interfering RNAs (siRNA) targeting VEGF-C could suppress lymphangiogenesis and lymph node metastasis in vivo. A lentivirus-based VEGF-C siRNA vector was infected into breast cancer cells and a xenograft model. The expression of VEGF-C mRNA and protein were quantified by quantitative real-time polymerase chain reaction (QRT-PCR), immunohistochemistry, and western blot analysis. The effect of VEGF-C siRNA on breast cancer cells was investigated by an invasion assay. Lymphangiogenesis was analyzed with anti-LYVE-1 and anti-D2-40 by immunohistochemical analysis. Lentivirus-mediated VEGF-C siRNA stably reduced VEGF-C mRNA And protein expression. VEGF-C siRNA inhibited the invasive ability of breast cancer cells in. vitro. Five weeks after intratumoral injection, the tumor volume was significantly smaller in the VEGF-C siRNA group than in the control scramble siRNA group in the MDA-MB-231 cell xenograft model. The numbers of LYVE-1 and D2-40 positive vessels per microscopic field were significantly decreased in the VEGF-C siRNA group, which indicates that VEGF-C siRNA inhibited lymphangiogenesis. Moreover, lymph node metastasis was significantly suppressed by VEGF-C siRNA in vivo. In conclusion, these results indicate that lentivirus-mediated VEGF-C siRNA offers a new approach for therapeutic intervention to prevent tumor growth and lymphatic metastasis of breast cancer. Anat Rec, 292:633-639, 2009. (C) 2009 Wiley-Liss, Inc.