siRNA抑制人舌癌细胞VEGF表达对MMP-2、-9及TIMP-1、-2蛋白表达的影响  

Effect of VEGF-siRNA on the expression of MMP-2,-9 and TIMP-1,-2 in human tongue squamous cell carcinoma (Tca8113) cell

作　　者：于大海[1] 郝洁[1] 李敬[1] 曹莹[1] 陈海波[1] 

机构地区：[1]广西医科大学附属口腔医院口腔颌面外科,南宁530021

出　　处：《北京口腔医学》2010年第1期1-4,共4页Beijing Journal of Stomatology

基　　金：国家自然科学基金资助项目(30360111)

摘　　要：目的利用小干扰RNA(small interfering RNA,siRNA)抑制人舌癌Tca8113细胞血管内皮生长因子(vascular endothelial growth factor,VEGF)表达,观察其对培养细胞和移植瘤基质金属蛋白酶(matrix metalloproteinases,MMPs)-2、-9及基质金属蛋白酶抑制剂(tissue inhibitors of metalloproteinases,TIMPs)-1、-2蛋白表达的影响。方法以稳定转染携带VEGF-siRNA真核表达载体的Tca8113细胞(VEGF-siRNA1、VEGF-siRNA2)为实验组,以转染空载体的及未转染的Tca8113细胞为实验对照组和空白对照组。将细胞分别接种于裸鼠皮下,用免疫组化法分别测定各组培养细胞和移植瘤VEGF、MMP-2、-9及TIMP-1、-2的蛋白表达。结果各组培养细胞及移植瘤VEGF免疫组化染色:VEGF-siRNA1组、VEGF-siRNA2组与实验和空白对照组相比,平均阳性率降低,平均灰度值增高(P<0.05);各组培养细胞均未检测出MMP-2、-9及TIMP-1、-2蛋白的阳性表达。各组移植瘤MMP-9、TIMP-1、-2可见阳性表达,但各组染色平均阳性率及平均灰度值之间的差异无统计学意义(P>0.05),而MMP-2未见明显阳性表达。结论以siRNA干扰沉默人舌癌Tca8113细胞VEGF基因,可明显抑制细胞及移植瘤内VEGF蛋白表达,但是对MMP-2、-9及TIMP-1、-2蛋白的表达无明显影响。Objective To investigate the expressions of matrix metalloproteinase （MMP） -2 ,-9 and tissue inhibitors of metalloproteinase （T IMP） -1,-2 in human tongue squamous cell carcinoma （Tca8113） in virto and in vivo. Methods The eukaryotic expression vector taken as an experiment control and two siRNA targeting VEGF constructed in the vectors （VEGF-siRNA1, VEGF-siRNA2 ） were transfected into human tongue squamous cell carcinoma cell lines（Tca8113）. The non-transfected Tca8113 cells were used as negative control. The transfected cells and non- transfected cells were injected subcutaneously on the back of the twenty nude-mice. The mice were executed after 47days of transfection. The expressions of VEGF,MMP-2 ,-9 and TIMP-1 ,-2 in the cultured cells and xenograft tumors were detected by immunocytochemistry and immunohistochemistry, respectively. Results Compared with the experimental and negative controls, the expressions of VEGF were decreased （ P 〈 0. 05 ） in two experimental groups of the cultured cells and the xenograft tumors. There were no obvious staining of MMP-2,-9 and TIMP-1,-2 in cultured cells and MMP-2 in the xenografts tissue in the four groups. There were no significant differences of the expressions of MMP-2 ,-9 and TIMP-1 ,-2 among the four groups of xenograft tumors （ P 〉 O. 05 ）. Conclusion VEGF-siRNA can down-regulate the expression of VEGF in the human tongue squamous cell carcinoma in vitro and in vivo. However,it may not influence the expressions of MMP-2,-9 and TIMP-1 ,-2.

关 键 词：小干扰RNA 舌癌 基质金属蛋白酶 基质金属蛋白酶抑制剂 血管内皮生长因子 

分 类 号：R739.8[医药卫生—肿瘤]