【摘要】 目的:研究TIMP-2基因转染对人成釉细胞瘤裸鼠移植瘤侵袭性生长的抑制作用。方法:实验分为空白对照组(Opti-MEM)、脂质体转染组(脂质体+Opti-MEM)和质粒转染组[pcDNA3.1(+)/GFP-TIMP-2+脂质体+Opti-MEM]。将以上各组液体混合注射于移植瘤的周围,每3天1次,共3次。采用RT-PCR分析移植瘤MMP-2及TIMP-2mRNA的表达,Western印迹分析移植瘤MMP-2及TIMP-2蛋白的表达,测定移植瘤体积和瘤重。应用SPSS11.0软件包对所得数据进行统计学分析。结果:接种AM组织块后,裸鼠生长良好,所有裸鼠在实验终止前均存活。接种AM组织块后1周末,可见裸鼠的接种处皮下有小结节,颜色与皮肤颜色相近,随皮肤移动而移动;2周末,小结节稍有增大,并且固定于皮下,不随皮肤的移动而移动,小结节表面较第1周略圆;实验终止时,所有移植瘤都有不同程度的增大。与对照组比较,质粒转染组移植瘤的生长速度在接种后4周明显降低。接种后第5周末,质粒转染组移植瘤的生长速度略有增加。自第1周末开始至实验终止时,质粒转染组移植瘤的体积均小于对照组。各组MMP-2mRNA表达平均值无变化,三者之间无显著性差异,P>0.05。质粒转染组的TIMP-2mRNA表达水平与空白对照组和空脂质体转染组比较均显著增加(P<0.05)。与空白组比较,质粒转染组的TIMP-2mRNA表达相对增加率为35.72%。质粒转染组MMP-2表达平均值比值为0.38,MMP-2蛋白抑制率为47.36%。质粒转染组MMP-2表达显著低于空白组,P<0.05。质粒转染组TIMP-2表达平均值比值为0.86,TIMP-2蛋白增加率为54.65%。质粒转染组TIMP-2表达显著高于空白组,P<0.05。结论:AM移植瘤的生长抑制,可能是由于TIMP-2基因过表达后,特异性抑制MMP-2蛋白所致。更多还原

【Abstract】 PURPOSE: To explore the invasiveness of AM xenografts in nude mice after TIMP-2 gene transfection. METHODS: Fresh ameloblastoma tissues were subcutaneously transplanted into the back of the nude mice.TIMP-2 expression plasmid was injected around the xenografts. The volume of the xenografts and the weight of xenografts in the terminal time of the experiment were recorded. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed to check the expression of MMP-2 and TIMP-2 mRNA in xenografts. Western blotting analysis was performed to investigate MMP-2 and TIMP-2 protein in xenografts. Students’ t test was used for statistical analysis with SPSS11.0 software package. RESULTS: All of the xenografts were survival. The volume of xenografts in control side wassignificantly larger than that in the experimental side. So did the weight of xenografts, P<0.05. The expression of TIMP-2 mRNA and protein in the experimental group was significantly higher than that in the control group. The expression of MMP-2 protein in experimental group was respectively significant lower than that in control group. CONCLUSION: The invasiveness and growth of xenograft of ameloblastoma was suppressed due to TIMP-2 gene transfection.更多还原