siRNA抑制人舌癌细胞VEGF表达对MMP-9蛋白表达的影响

The expression of MMP-9 after the expression of VEGF suppressed by siRNA in human tongue squamous cell carcinoma

目的利用小干扰RNA（small interfering RNA，siRNA）抑制人舌癌Tca883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-9（matrix metalloproteinase-9，MMP-9）蛋白表达的影响。方法以稳定转染携带VEGF—siRNA真核表达载体的人舌癌Tca8113细胞（VEGF—siRNA1、VEGF—siRNA2）为实验组，以转染空载体人舌癌Tca8113细胞及未转染的人舌癌Tea8113细胞为实验对照组和空白对照组，将各组细胞分别接种于裸鼠皮下，用免疫细胞／组织化学法分别测定各组培养细胞和移植瘤VEGF及MMP-9的蛋白表达。结果在培养细胞和移植瘤中，与实验对照组和空白对照组相比，VEGF—siRNA1组和VEGF—siRNA2组VEGF染色平均灰度值高，差异有统计学意义（P〈0．05）；各组培养细胞中均未见MMP-9表达；各组移植瘤MMP-9染色平均灰度值之间的差异无统计学意义（P〉0．05）。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-9蛋白的表达无明显影响。

Objective To investigate the expression of matrix metalloproteinase -9 （MMP -9） in human tongue squamous cell carcinoma（ Tca8113 ）in vitro and in vivo, after the expression of vascular endothelial growth fac- tor（VEGF） suppressed by vector- based small interfering RNA （siRNA）. Methods The eukaryotic expression vec- tor taken as an experimental control and two siRNA targeting VEGF constructed in the vector （ 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 injec- ted subcutaneously on the back of the twenty nude - mice which were divided into four groups randomly （ n -- 5/ group）, respectively. The expression of VEGF and MMP - 9 in the cultured ceils and the xenograft tumors were de- tected by immunocytochemistry and immunohistochemistry. Results Compared with the experimental and negative controls, the VEGF expressions were decreased （P 〈 0.05 ） in two experimental groups of the cultured cells and xen- ograft tumors. There was no obvious MMP - 9 staining in four groups of cultured cells. There were no significant differences of the MMP - 9 expression among four groups of xenograft tumor （ P 〉 0. 05 ）. Conclusion VEGF - siR- NA can down - regulate the expression of VEGF in the human tongue squamous cell carcinoma in vitro and in vivo. However, it may not influence on the expression of MMP -9 efficiently.