【摘要】 目的　建立检测组织因子 (tissuefactor ,TF)mRNA荧光定量的方法 ,了解组织中TF的表达水平 ,研究TFmRNA的表达与肿瘤的关系。方法　构建克隆载体 pUC TF作为定量模板 ,在GeneAmp 5 70 0型检测仪上定量检测 4 2例不同恶性程度胶质瘤组织标本 ,5例正常脑组织标本和人恶性胶质瘤细胞株U 2 5 1。结果　 4 2例肿瘤组织均有mRNA的表达 ,范围从 3 .6× 10 5～ 8.8× 10 7拷贝 / μgRNA ,均值为 2 .9× 10 6 拷贝 / μgRNA ,且拷贝数与恶性程度成正比 ;5例正常脑组织中 ,1例检测不出 ,其他 4例的强度为 3 .8× 10 3～ 5 .1× 10 4拷贝 / μgRNA ,均值为 6.2× 10 3拷贝 / μgRNA ;胶质瘤细胞株U 2 5 1的强度为 2 .8× 10 6 拷贝 / μgRNA。 结论　成功地建立了荧光定量检测TF基因表达方法 ,检测结果可用绝对拷贝数表示 ,定量准确可靠。TFmRNA荧光定量测定方法的建立为研究TF与胶质瘤等肿瘤恶性程度及转移的关系打下了基础更多还原

【Abstract】 Purpose To establish a FQ?PCR (fluorogenic quantitative RT?PCR) method for detecting the expression of tissue factor gene in tissue and investigate the relationship between TF mRNA expression and tumor. Methods Fluorogenic quantitative RT?PCR was set up to examining the specific expression of tissue factor in 42 glioma tissues,5 normal brain tissue and glioma cell line U?251. Results In 42 glioma tissues TF mRNA expression ranges from 3.6×10 5-8.8×10 7 copy/μg RNA,mean value is 2.9×10 6 copy/μg RNA.TF mRNA expression level correlates with the histologic grade of malignancy.In one of 5 normal brain specimens,TF mRNA is under detection limit whereas in the other 4 normal specimens TF mRNA expression level is from 3.8×10 3-5.1×10 4 copy/μg RNA,mean value is 6.2×10 3 copy/μg RNA.For glioma cell line U?251 the expression level is 2.8×10 6 copy/μg RNA. Conclusions Tissue factor gene expression detected with FQ?PCR had an accurate,speedy results.TF mRNA may be used as a valuable marker for evaluating the grade of glioma malignancy.更多还原