粒细胞弹性蛋白酶表达与肾癌预后的关系

目的: 探讨肾癌多型核粒细胞弹性蛋白酶的表达与肾癌预后的关系.方法: 应用酶联免疫吸附实验研究肾癌组织中多型核粒细胞弹性蛋白酶的表达;病理切片CD34染色,检测肾癌细胞对微血管的侵袭;检测肾癌多型核粒细胞弹性蛋白酶(polymorphonuclear leukocyte elastase ,PMNE)启动子基因重复序列突变,应用pGL-3 Basic载体分别构建突变型及野生型PMNE荧光素酶真核表达载体(pGL/PMNE), 分别测定荧光素酶活性,探讨多型核粒细胞弹性蛋白酶启动子重复序列突变的意义.结果: 59例肾癌组织中多型核粒细胞弹性蛋白酶的表达明显高于22例正常肾组织(P<0.01);肾癌组织中多型核粒细胞弹性蛋白酶的表达与肿瘤Fuhrmans分期密切相关(P<0.01);肾癌组织中癌细胞对微血管的侵袭与多型核粒细胞弹性蛋白酶的表达关系密切(P<0.01);59例肾癌组织中发现1例多型核粒细胞弹性蛋白酶启动子重复序列突变:-879/-716TCCA,荧光素酶活性试验显示该突变对其下游基因表达有明显增强作用.结论: 多型核粒细胞弹性蛋白酶的表达与肾癌的临床病理学表现关系密切,多型核粒细胞弹性蛋白酶启动子重复序列-879/-716TCCA突变可能与肾癌PMNE的表达及侵袭能力有关.多型核粒细胞弹性蛋白酶在肾癌的发展过程中具有重要的作用.

Relationship between expression of polymorphonuclear leukocyte elastase and renal cell carcinoma

AIM: To study the relationship between the expression of polymorphonuclear leukocyte elastase (PMNE) and renal cell carcinoma (RCC) patients. METHODS: By molecular, immunological and pathological methods, we measured the PMNE expression in the cytoplasmic extracts of 59 renal cell carcinomas, observed the microvascular invasion of RCC cells and sequenced the PMNE promoter gene of RCC. The result of promoter sequencing showed a nucleic acid mutant. The mutated promoter activity was also compared with that of wild type by constructing pGL3/PMNE luciferase expression vector. RESULTS: The PMNE was over expressed in RCC cells (P<0.01) and the expression levels of PMNE in the tissue extracts of G3 and G4 were much higher than those in the tissue extracts of G1 and G2(P<0.01). The PMNE over-expression was also detected in the specimen of RCC with microvascular invasion(P<0.01). In one case, permutation mutant from A to C in the-879/-716 (TCAA to TCCA) was found. The luciferase expression in K562 cells transfected with mutated PMNE promoter luciferase expression vector was markedly higher than those transfected with wild type PMNE promoter vector. CONCLUSION: PMNE is over expressed in RCC and the level of its expression is closely related with the stage and the invasion ability of the tumor. PMNE promoter gene permutation mutant, -879/-716TCAA to TCCA, may lead to higher PMNE enzyme activity, which promotes the invasive potential of RCC cells. Our data indicate that PMNE may play an important role in the development of RCC.