成釉细胞瘤中MMP-2的表达及RNA干扰的抑制作用  

Expression of MMP-2 in ameloblastoma cells and inhibition by RNA interference

作　　者：曾东林[1] 黄洪章[1] 张彬[1] 潘朝斌[1] 梁蔚文[1] 

机构地区：[1]中山大学附属第二医院口腔颌面外科,广东广州510120

出　　处：《中国口腔颌面外科杂志》2005年第4期315-319,共5页China Journal of Oral and Maxillofacial Surgery

基　　金：国家自然科学基金(30471896);广东省自然科学基金(04300340)

摘　　要：目的:研究MMP-2在成釉细胞瘤中的表达和RNA干扰对成釉细胞瘤细胞中MMP-2基因的沉默效应。方法:培养成釉细胞瘤细胞,应用免疫组织化学方法检测成釉细胞瘤中MMP-2的表达;体外构建MMP-2靶向siRNA质粒表达载体,将siRNA质粒表达载体转染原代培养的成釉细胞瘤细胞,激光共聚焦显微镜检测siRNA质粒表达载体的转染效率,RT-PCR检测成釉细胞瘤细胞中MMP-2mRNA的改变,明胶酶谱法检测MMP-2的变化,应用SPSS11.0统计软件中的t检验对结果进行统计学分析。结果:成釉细胞瘤中MMP-2呈阳性表达,siRNA质粒表达载体对原代培养的成釉细胞瘤细胞的转染率为41.8%;转染后,成釉细胞瘤细胞的MMP-2mRNA表达水平显著降低,酶原性MMP-2和活性MMP-2显著减少,P<0.05。结论:体外构建的MMP-2靶向siRNA质粒表达载体可以转染体外培养的成釉细胞瘤细胞,并导致MMP-2基因沉默。PURPOSE： To study the expression of MMP-2 in ameloblastoma and the inhibition of MMP-2 gene by RNA interference. METHODS： Primary ameloblastoma cells cultured in Defined Keratinocyte-SFM medium in vitro and MMP-2 detected by immunohistoehemisty. Plasmid-vector for siRNA was eonstrueted and transfeeted into ameloblastoma cells. Laser confoeal microscope was used to detect the efficiency of transfeetion. Reverse transcription polymerase chain reaction analysis was performed to ehcek the expression of MMP-2 mRNA. Gelatinolytic zymography analysis was used to investigate the activity of MMP-2. Student t test was used to analyze the data. RESULTS： Positive expression of MMP-2 was observed in ameloblastoma. The transfection rate of plasmid-vector for siRNA into ameloblastoma cells was 41.8%. The expression level of MMP-2 mRNA and pro-MMP-2 and active MMP-2 were significantly decreased after plasmidbased siRNA transfeetion （P〈0.05）. CONCLUSION： MMP-2 targeted plasmid-vector for siRNA constructed in vitro can be successfully transfected into ameloblastoma cells cultured in vitro and can silence MMP-2 gene.

关 键 词：MMP-2 RNA干扰 转染 siRNA质粒表达载体 基因沉默 抑制作用 成釉细胞瘤 

分 类 号：R739.8[医药卫生—肿瘤]