实时荧光定量RT - PCR 法检测膀胱移行细胞癌VEGF的表达和意义.DOCVIP

临床医学论文-实时荧光定量RT - PCR 法检测膀胱移行细胞癌VEGF的表达和意义 ??????????? 作者：安全意，张金刚，刘德斌，李成，马可为，姜丽萍 【摘要】? 目的 建立实时荧光定量逆转录-多聚酶链反应（real-time quantitative, RT-PCR）检测膀胱移行细胞癌（BTCC）组织VEGF mRNA表达的方法,并验证其与免疫组织化学检测VEGF蛋白表达的相关性。方法 采用免疫组织化学SP法和实时荧光定量RT-PCR法分别从蛋白水平和mRNA水平检测了31例BTCC和15例正常膀胱黏膜组织中VEGF的表达。结果 BTCC中VEGF mRNA表达水平显著高于正常组，两组比较差异有显著性（P0.05）;BTCC中表达水平在高分化组与中低分化组之间、Tis-T1期与T2-T4期间的差异均有显著性（P0.05）。VEGF的阳性表达率在BTCC组中为100%,显著高于正常组（P0.05），在高分化组与中低分化组之间、Tis-T1期与T2-T4期间的差异均有显著性（P0.05）。结论 本研究所建立的用于检测人BTCC组织VEGF mRNA表达的实时荧光定量RT-PCR方法,是一种快速有效、灵敏度高、特异性好的定量检测方法,与免疫组化SP方法检测的VEGF蛋白表达具有良好相关性，可作为研究膀胱肿瘤的生物学行为和判断预后的参考指标。 【关键词】? 膀胱癌;血管内皮生长因子;免疫组织化学；实时荧光定量RT-PCR ??? 【Abstract】? Objective? To establish a real-time fluorescence quantitative RT-PCR for detection of VEGF mRNA expression in bladder transitional cell carcinoma （BTCC） and verify the correlation with protein expression detected by the immunohisochemistry method.Methods? Total RNA was extracted with Trizol from the tissues and mRNA was transcribed reversely into cDNA. The real-time quan-titative RT-PCR was used to detect the expression of VEGF mRNA in 31 in BTCC and 15 control normal bladder mucosa tissues. The immunohistochemistry SP method was used to evaluate the protein expression of VEGF.Results? The expression of VEGF mRNA was significantly higher in BTCC tissues than that in the normal bladder mucosa tissue tissues （P0.05）. BTCC expression levels in well-differentiated group and between groups of poorly differentiated, Tis-T1 with T2-T4 period differences were statistically significant （P0.05）.Conclusion? Real-time fluorescence quantitative RT-PCR used to detect the expression of VEGF mRNA might be a reliable method for early diagnosis of BTCC. The expression of VEGF mRNA is correlated with that of the VEGF protein significantly. It can be used to study the biological behavior of bladder cancer and judg the prognosis. ??? 【Key words】? bladder cancer;vascular endothelial growth factor;immunohistochemistry;real-time fluorescence quantitative RT-PCR