【作者】 李守淼；

【导师】 王家祥； 范应中；

【作者基本信息】 郑州大学 ， 外科学， 2004， 硕士

【摘要】 背景与目的 肾母细胞瘤(Nephroblastoma)又称肾胚胎瘤，是小儿最常见的腹部恶性实体瘤，占肾脏肿瘤第一位(97％)。Max Wilms于1899年对该瘤的特征进行了详细的描述，故被命名为Wilms’瘤。肾母细胞瘤是由胚胎性肾组织发生的一种恶性混合瘤，常发生局部浸润和淋巴、血运转移，多发生于6个月至3岁的婴幼儿期。该肿瘤虽为恶性，但目前认为它是一种可治性疾病，预后好于其它恶性肿瘤。组织学分型和临床分期仍是主要的制定治疗方案和预测预后的标准。尚未发现特异的肿瘤诊断分子标记物。现已证实，间变型是独立的预后因素，预示预后不良。 uPA(urokinase-type plasminogen activator，uPA)及其受体(uPA receptor，uPAR)和抑制剂(plasminogen activator inhibitor-1，PAI-1)属于尿激酶型纤溶酶原激活剂系统(uPA system，uPAs)，在降解细胞外基质、溶解基底膜过程中起重要作用。这三者间相互作用，共同调节着肿瘤细胞的浸润、血管生成和转移。通过研究三者在肾母细胞瘤浸润、转移中的作用，可为其将来的生物学治疗提供可能的途径。关于上述三种蛋白在肾母细胞瘤中的表达，目前国内外尚未见报道。本文旨在探讨这三种蛋白在肾母细胞瘤发生、发展、临床诊断及预后判别中的价值，为肾母细胞瘤的基因治疗提供一种富有前景的新方向。郑州大学2004年硕士研究生毕业论文uP人及其受体(uPAR)和抑制剂(PA卜1)在肾母细 胞瘤中的表达及意义(中文摘要) 本研究采用免疫组化技术，对30例肾母细胞瘤和标本中uPA、uPAR和PAI一1蛋白进行检测，以探讨15例非肿瘤肾组织石蜡uPA、uPAR和白在肾母细胞瘤中的表达情况及其与组织学分型、临床分期的关系，三者在肿瘤的浸润、转移、血管生成及预后中的作用。PAI一1蛋借以了解材料与方法 本研究收集了手术切除的肾母细胞瘤石蜡标本30例，术前均未放化疗，其中男19例，女n例，年龄7月~9岁，中位年龄2.6岁，按美国肾母细胞瘤研究组(National Wilms，Tumor stody oro即，NwTs一5)标准[2]进行分组，组织学分型分为预后良好组织型(favorable histology，FH)和预后不良组织型(unfavorable histology，UH);l隋床分期分为低分期组和高分期组。其中，FH型23例，UH型7例;低分期组13例，高分期组17例。对照组15例为肿瘤旁正常肾组织或因非肿瘤切除的肾组织石蜡标本。其中，肿瘤旁肾组织6例，非肿瘤肾组织9例。每例均作HE染色和免疫组化染色。采用免疫组化SABC法检测uPA、uPAR和PAI一1蛋白的表达情况。所有资料采用SPSS 10.0软件处理，统计数据用x士，表示，统计处理采用用犷检验、Fishe;’，精确概率检验和t检验，uPA、uPAR和PAI一1之间的关系行直线相关分析，以a二0.05为检验水准。结果 1.uPA在肾母细胞瘤组织中阳性率为66.7%(20/30)，而在对照组中仅有1例阳性，差别具有显著性(尸<0.05)。在对照组标本中，部分肾小管呈淡黄色，极少数血管内皮细胞呈黄色或棕黄色染色。uPA在肾母细胞瘤组织中的表达与组织学分型、临床分期显著相关，在UH型和临床高分期组中的表达明显强于FH型和低分期组，差异具有显著性(P<0.05)。 2.uPAR在肾母细胞瘤组织中阳性率为56.7%(17/30)，而在对照组中均为阴性，差别具有显著性(P<0.05)。在对照组中仅有部分肾小管呈淡黄色，极少数血管内皮细胞呈黄色或棕黄色染色。uPAR在肾母细胞瘤组织中的郑州大学2004年硕士研究生毕业论文uPA及其受体〔uPAR)和抑制剂(PA卜1)在肾母细 胞瘤中的表达及意义(中文摘要)茄;宁妙二然厂麒毛健男诵扩隽毅护滋梦余费诺魏添撇犷谕鱿、泌男支考表达也与组织学分型、临床分期显著相关，在FH型和临床低分期组中的表达较UH型和高分期组弱，差异具有显著性(P<0.05)。3.PAI一1在肾母细胞瘤组织中阳性率为50.0%(15/30)也均为阴性，强阳性表达;差别具有显著性(尸<0.05)。肿瘤组中，PAI.1，而在对照组中在肿瘤细胞上呈而在对照组中，仅有部分肾小管呈淡黄色，极少数血管内皮细胞呈黄色或棕黄色染色。PAI一1在肾母细胞瘤组织中的表达与组织学分型、临床分期显著相关，差异具有显著性(P<0.05)。4.在肾母细胞瘤组织中uPA、uPAR和PAI一1三者的表达具有明显直线正相关(P<0，05)。结论 1.uPA、uPAR和PAI一1蛋白在肾母细胞瘤中有较强表达，而在非肿瘤肾组织中仅有弱表达或无表达。而且在肿瘤组中的表达与组织学分型和临床分期显著相关。说明三者在肾母细胞瘤的浸润转移和血管生成中发挥一定作用。2.uPA、uPAR和PAI一1三者的表达之间呈明显正相关，说明三种蛋白同时参与了肿瘤的浸润和转移。3.可以根据三者的表达情况判断转移和局部复发的风险，以便采取积极的综合治疗，提高患者的存活率。4.uPA、uPAR和PAI一1可以作为肾母细胞瘤基因治疗的一组靶点，本研究为临床合理有效治疗肾母细胞瘤提供了一种新思路。更多还原

【Abstract】 Background and ObjectiveNephroblastoma, or named embryonic tumor of kidney, is one of the most common pediatric abdominal malignant tumors in children. It consists of 97% in all renal tumors. In 1899, Max Wilms described its characteristics, so it was named after his name. Nephroblastoma is malignant mixed tumor, derived from embryonic renal tissue. It always occurred local spread, lymphatic metastasis and hematogeneous metastasis in 6 mouths to 3 years. At present, it is considered that nephroblastoma was a curable disease, and its prognosis was better than other malignant tumors, although it was malignant. Histological type and clinical stage are important criteria in instituting heal project and prediction of outcome. An independent prognostic molecular marker has not been identified as yet. Anaplasia type has been proven to be of prognostic value and is associated with poor prognosis.Urokinase-type plasminogen activator (uPA), its receptor (uPAR) and plasminogen activator inhibitor- 1(PAI-1) are part of the urokinase-type plasminogen activator system, playing an important role in degrading extracellular matrix and dissolution of the basement membrane. These proteins regulate together infiltration of tumor cell, angiogenesis and metastasis. Via studying theirexpressions in nephroblastoma, we will provide a possible way in its biological therapy. There was no report about these proteins’ expression in nephroblastoma in our internal periodical. The purpose of this text is to study the proteins’ value of occurrence, development, clinical diagnosis and forecasting prognosis in nephroblastoma, and provide a new direction of full of foreground for gene therapy of nephroblastoma.In this research, the expression of uPA, uPAR and PAI-1 proteins was detected by the immunohistochemistory SABC method in 30 nephroblastoma specimens (tumor group) and 15 non-neoplasia renal tissue specimens (control group), and all the sections were formalin-fixed and paraffin-enbeded. The purpose of this research is to study expressions of three proteins and relations with histological types and clinical stages and to find out those proteins’ roles in tumor’s local spread, metastasis, angiogenesis and prognosis.Materials and MethodsThirty nephroblastoma specimens had been collected from surgical resections. These patients, from 7 months to 9 years, average 2.6 years, 19 males and 11 females, were not performed any radiotherapy and chemotherapy. By NWTS-5 criteria, histological types of their HE staining sections were classified by two types. Among them 23 patients belonged to favorable histology (FH) type, and 7 patients did unfavorable histology (UH) type. Clinical stages were classified by two types. Among them 13 patients belonged to low clinical stage, and 17 patients did high clinical stage. Control group, 15 patients, consisted of 6 normal renal tissues near tumor and 9 non-neoplasia renal tissues. Every specimen had been stained in HE staining and immunohistochemistory. Expression of uPA, uPAR and PAI-1 patients was detected by SABC method. The results were analyzed by SPSS 10.0 statistical software. Data were presented as mean ?SEM. Statistical analysis was done by Pearson’s chi-square test, Fisher’s exact test or Student’s t test. The relationships between expression of uPA, uPAR, and PAI-1 were evaluated by the linear correlation test. =0.05 were considered significant test level.Results1. The positive rate of uPA in nephroblastoma group was 66.7% (20/30), but in control group the positive rate was only 6.7%(1/15). There was significant difference (P <0.05). In control group, a part of renal tubules had weak staining and a few of vessel endotheliocytes showed a very intense staining. Significant correlateions were caught between expression and histological type or beteeen eppression and clinical stage. There were significant difference among them(P <0.05). In UH type and in high clinical stage the expression was stronger than in FH type and in low clinical stage.2. The positive rate of uPAR in nephroblasto更多还原