原代培养人喉鳞癌细胞中SIX1、TGF-β、VEGF-C表达的相关性  

Correlation Between Expression of SIX1, TGF-β and VEGF-C in Primarily Cultured Human Laryngeal Squamous Cell Carcinoma Cells

作　　者：张继华[1] 陈春菊[1] 韩彩莉 苏静[1] 李丹[1] 王来[2] 

机构地区：[1]河北医科大学第一医院耳鼻咽喉科,石家庄050031 [2]河北医科大学第一医院重症医学科,石家庄050031

出　　处：《肿瘤防治研究》2016年第6期453-458,共6页Cancer Research on Prevention and Treatment

摘　　要：目的探讨原代人喉鳞癌细胞中SIX1、TGF-β、VEGF-C表达相关性。方法取新鲜人喉鳞状细胞癌组织,进行原代细胞培养,利用基因干扰技术,制备SIX1、TGF-β及SIX1+TGF-β靶向si RNA转染至人喉鳞癌细胞,转染成功后将实验细胞分为5组:A组(未转染组)、B组(转染空载体组)、C组(SIX1-si RNA组)、D组(TGFβ-si RNA组)、E组(SIX1+TGF-β-si RNA组)。Western blot及细胞爬片免疫荧光检测各组细胞SIX1、TGF-β及VEGF-C蛋白的表达。RT-PCR检测各组细胞SIX1、TGF-β及VEGF-C m RNA的表达。结果 SIX1、TGF-β、VEGF-C蛋白及其m RNA的表达在未转染组、转染空载体组中差异无统计学意义。同未转染组相比,SIX1蛋白及其m RNA的表达在SIX1-si RNA组降低的同时出现了VEGF-C表达的降低;TGF-β蛋白及其m RNA的表达在TGFβ-si RNA组降低的同时也出现了VEGF-C表达的降低,差异均有统计学意义。同SIX1-si RNA组、TGFβ-si RNA组相比,VEGF-C蛋白及其m RNA的表达在SIX1+TGF-β-si RNA组未出现进一步降低。结论 SIX1和TGF-β能够共同作用促进VEGF-C蛋白及其m RNA的表达,进而促进人喉鳞癌的淋巴转移。SIX1、TGF-β的变化可能成为临床抑制人喉鳞癌淋巴结转移的潜在靶点。Objective To investigate the relationship between sine oculis homeobox homolog 1（SIX1）, TGF-β and VEGF-C in primarily cultured human laryngeal squamous cell carcinoma cells. Methods We took fresh human laryngeal squamous cell carcinoma tissues and cultured the primary cells. The technology of RNA interference was used for preparing SIX1-targeting, TGF-β-targeting and SIX1＋TGF-β-targeting si RNA to transfect laryngeal squamous cell carcinoma cells. After successful transfection, the experiments were divided into five groups： Group A（untransfected）, Group B（empty vector）, Group C（SIX1-si RNA）, Group D（TGF-β-si RNA） and Group E（SIX1＋TGF-β-si RNA）. The protein expression of SIX1, TGF-β and VEGF-C were determined by immunohistochemistry and Western blot. The m RNA levels of SIX1, TGF-β and VEGF-C were determined by RT-PCR. Results The protein and m RNA expression of SIX1, TGF-β and VEGF-C in empty vector group had no significant difference compared with untransfected group. In SIX1-si RNA group, the expression of SIX1 protein and m RNA were decreased, at the same time the expression of VEGF-C was decreased, with statistically significant difference compared with untransfected group. In TGF-β-si RNA group, the expression of TGF-β protein and m RNA were decreased, at the same time the expression of VEGF-C was decreased, with statistically significant difference compared with untransfected group. Compared with SIX1-si RNA and TGF-β-si RNA groups, the expression of VEGF-C protein and m RNA had no significant decrease in SIX1＋TGF-β-si RNA group. Conclusion SIX1 could promote lymph node metastasis by coordinating with TGF-β to increase the expression of VEGF-C protein and m RNA, suggesting that SIX1 and TGF-β might be potential therapeutic targets for preventing lymph node metastasis from human laryngeal squamous cell carcinoma.

关 键 词：SIX1 TGF-Β VEGF-C 原代人喉鳞癌细胞 

分 类 号：R737.33[医药卫生—肿瘤]