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

siRNA抑制人舌癌细胞VEGF表达对MMP-2、MMP-9及
TIMP-1蛋白表达的影响
陈凤强;郝洁;于大海;李晶;罗婷婷
【期刊名称】《广西医科大学学报》
【年(卷),期】2012(029)004
【摘要】Objective: To investigate the effects of vascular endothelial growth factor (VEGF) suppression by vector-based small interfering RNA (siRNA) on the expression of matrix metalloproteinase (MMP)-2, MMP-9 and tissue inhibitors of metalloproteinase-1 (TIMP-1) in human tongue squamous cell carcinoma (Tca8113) in vitro and in vivo. Methods: The eukaryotic expression vector was taken as an experimental control 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 subcutane-ously on the back of twenty nudemice which were divided into four groups randomly ( n
=5/group) ; respectively. The expression of MMP-2, MMP-9 and TIMP-1 in xenograft tumors -was detected by Gelatin zymography and Western blot protein immunoblotting. Results: Compared -with the experimental and negative controls, the VEGF expression was decreased ( P <0. 05) in two experimental groups of the xenograft tumors. There -was no significant difference in the expression of MMP-2, MMP-9 and TIMP-1 among four groups of xenograft tumor ( P >0. 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 affect the expression of MMP-2, MMP-9 and TIMP-1 efficiently.%目的:探讨小干扰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)表达对移植瘤基质金属蛋白酶(matrix metalloproteinases,MMP)-2、MMP-9及基质金属蛋白酶抑制剂-1(tissue inhibitors of metalloproteinase-1,TIMP-1)蛋白表达的影响.方法:以稳定转染携带针对VEGF的siRNA真核表达载体的人舌癌Tca8113细胞(VEGF-siRNA1、VEGF-siRNA2)为实验组,以转染空载体的人舌癌Tca8113细胞及未转染的人舌癌Tca8113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,用明胶酶谱实验及蛋白免疫印迹法分别测定各组移植瘤TIMP-2、TIMP-9及TIMP-1的蛋白表达.结果:各组明胶酶谱实验结果均有较明显MMP-9蛋白表达,活性MMP-9蛋白含量较少,MMP-2蛋白表达量明显少于MMP-9蛋白表达,而活性MMP-2几乎完全没有可见条带出现.各组中MMP-2、MMP-9及活性-MMP-9条带酶量差异均尚不具统计学意义(P＞0.05).以scion图像分析系统分别读取各样本TIMP-1条带灰度值及β-actin条带灰度值,各组中TIMP-1蛋白相对含量差异亦无统计学意义(P＞0.05).结论:以siRNA干扰人舌癌Tca8113细胞VEGF基因,可明显抑制肿瘤内VEGF的蛋白表达,但是对MMP-2、MMP-9及TIMP-1蛋白的表达无明显影响.
【总页数】3页(P493-495)
【作者】陈凤强;郝洁;于大海;李晶;罗婷婷
【作者单位】广西医科大学附属口腔医院口腔颌面外科,南宁,530021;广西医科大学附属口腔医院口腔颌面外科,南宁,530021;广西医科大学附属口腔医院口腔颌面
外科,南宁,530021;广西医科大学附属口腔医院口腔颌面外科,南宁,530021;广西医科大学附属口腔医院口腔颌面外科,南宁,530021
【正文语种】中文
【中图分类】R739.86
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