Department of Pathology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, China.
J Cancer Res Clin Oncol. 2012 Jan;138(1):133-9. doi: 10.1007/s00432-011-1079-9. Epub 2011 Nov 6.
We investigated the role of vascular endothelial growth factor C (VEGF-C) in esophageal squamous cell carcinoma (ESCC) by knocking down VEGF-C expression in the ESCC cell line EC9706.
Immunohistochemistry and in situ hybridization techniques were used to detect the expression of VEGF-C expression in ESCC tissues. We also investigated the relationship between VEGF-C expression and lymph node metastasis. We designed a siRNA expression plasmid for VEGF-C and transfected it into EC9706 cells. Stable clones were selected, and VEGF-C expression was analyzed by RT-PCR and western blotting. Cells were inoculated into nude mice. The expression of VEGF-C in the resulting tumors was analyzed by immunohistochemistry and in situ hybridization.
VEGF-C is highly expressed in ESCC and correlated with lymph node metastasis, as high levels were observed in patients presenting with lymph node metastases relative to those who did not (P < 0.01). Transfection with VEGF-C-siRNA decreased the expression of VEGF-C mRNA and protein. ESCC cells stably transfected with VEGF-C-siRNA expressed very low levels of VEGF-C (P < 0.01 compared with control). This knockdown effect persisted when the cells were inoculated into nude mice and allowed to form tumors.
The siRNA-targeted knockdown of VEGF-C led to a significant reduction in VEGF-C expression. This siRNA technique could be used for gene therapy in ESCC.
通过在 ESCC 细胞系 EC9706 中敲低 VEGF-C 的表达，研究血管内皮生长因子 C（VEGF-C）在食管鳞状细胞癌（ESCC）中的作用。
采用免疫组织化学和原位杂交技术检测 ESCC 组织中 VEGF-C 表达情况。还研究了 VEGF-C 表达与淋巴结转移之间的关系。我们设计了针对 VEGF-C 的 siRNA 表达质粒，并将其转染到 EC9706 细胞中。选择稳定的克隆，通过 RT-PCR 和 Western blot 分析 VEGF-C 的表达。将细胞接种到裸鼠中。通过免疫组织化学和原位杂交分析肿瘤中 VEGF-C 的表达。
VEGF-C 在 ESCC 中高表达，与淋巴结转移相关，有淋巴结转移的患者表达水平高于无淋巴结转移的患者（P<0.01）。转染 VEGF-C-siRNA 可降低 VEGF-C mRNA 和蛋白的表达。稳定转染 VEGF-C-siRNA 的 ESCC 细胞 VEGF-C 表达水平非常低（与对照组相比，P<0.01）。这种敲低效应在细胞接种到裸鼠并允许形成肿瘤时仍然存在。
针对 VEGF-C 的 siRNA 靶向敲低导致 VEGF-C 表达显著降低。这种 siRNA 技术可用于 ESCC 的基因治疗。
