【作者】 田俊严；

【导师】 杨启政；

【作者基本信息】 郑州大学 ， 小儿外科， 2002， 硕士

【摘要】 肾母细胞瘤(Nephroblastoma，Wilms’瘤)是小儿最常见的腹部恶性肿瘤之一，1899年德国医师Wilms首次报道此病，故命名为Wilms’瘤。因本瘤在胚胎发生上是由后肾胚基发展而来，且肿瘤由极其类似肾母细胞的成分所组成，故又称为肾母细胞瘤。在儿童恶性肿瘤中，其发病率在国外仅次于神经母细胞瘤，但在我国则略高于神经母细胞瘤，而居小儿腹部恶性肿瘤首位。肿瘤主要发生在出生后最初5年内，特别多见于2～4岁，常与先天畸形并发，如先天性虹膜缺如、先天性单侧肢体肥大等。肿瘤起源于未分化的肾胚基，可形成肾的各种成分，是间叶组织和上皮组织组成的恶性混合瘤。本瘤的预后与肿瘤的分化程度，发病年龄，转移与否，是否为双侧肿瘤及手术切除的彻底性等有关，至今尚无特异性肿瘤诊断标记物发现。 本研究采用免疫组织化学技术，探讨血管内皮生长因子郑州大学2002年硕士毕业论文 贤母细胞瘤中VEGF和NOS e与血管生成的关系及临床意义 （VEGF）内皮型一氧化氮合酶＊NOS）在肾母细胞瘤微血管生成中的作用，探讨它们之间的关系及其与肾母细胞瘤的发生、发展等生物学特性的相关性，为肾母细胞瘤治疗方案的设计和肿瘤的防治提供新的思路和依据。 材料与方洁：本实验收集手术切除的小儿肾母细胞瘤标本50例，标本均经10％福尔马林固定，石腊包埋。50例患者年龄最小为 4个月，最大的 15岁，平均年龄为 5．44士 4．73岁，其中男性为 32例，女性为 18例。患儿术前均未经放疗及化疗。HE切片明确诊断并作组织病理分类。根据术中情况及病理诊断判定临床病理分期（按NWTSl标准人 其中临床病理分期I～11期30例，Ill～N期为20例。组织病理分类：组织结构良好型吓H型）40例，组织结构不良型（UH型）10例。采用兔疫组织化学方法旧ABC法）检测全部标本中MVD、VEGF和 eNOS的表达情况。eNOS与 VEGF均以在肿瘤组织中细胞阳性数＜10％为阴性，＞10％为阳性。＊＊D结果判断以5个高血管密度区在高倍视野中的血管数目均值来表示。实验数据应用肥SSm刀软件包处理，P＜0刀5认为有统计学意义的检查标准。 结果：门）MVD值在肾母细胞瘤 FH型中为 5 0．5 7 fi 3．of，在 UH型中为 85．92上 14．36，H者相比有统计学意义。I～11期 MVD值为 49．14士 15．70，Ill～IV期为 70．39f 1759，二者 2 郑州大学 2002年硕士毕业论文 肾母细胞痈中 VEGF和 NOS 4ti！i与血管生成的关系及临床意义 相比也具有统计学意义。 Q）VEGF在‘肾母细胞瘤FH型中表达率为65．00O，在UH 型中表达率为60刀0％，二者相比无统计学意义。VEGF在l～ 11期表达率36石7％，Ill～N期表达率为80刀0％，二者相比有 统计学意义。 门eNOS在肾母细胞瘤FH型中表达率为27．50％，在 UH型中表达率为70刀0％，二者相比具有统计学意义；eNOS 在I～11期表达率为20．000，而在Ill～IV期表达率为60．00％， 二者相比也具有统计学意义。 N）MVD值在 VEGF阳性组中为 65．70士 16．88，阴性组 为 48．18118．13，二者相比有统计学意义；MVD值在 eNOS阳性 组中为刀．13t19．38，在阴性组中为48．93t15．09，二者相上有统 计学意义。 乃）VEGF及 eNOS均表达阳性 10例，均表达阴性 15 例。VEGF表达阳性而 eNOS表达阴性 17例，eNOS表达阳性 而VEGF表达阴性者8例。VEGF在肾母细胞瘤中表达与eNOS 表达之间无相关性。 “）正常对照组VEGF、eNOS均表达阴性。 结论：l、MVD值是判断肾母细胞瘤预后一较为理想的 指标，MVD值与组织病理分型及临床分期有明显相关性。2。 VEGF表达与其组织病理分型无关，而与；ha床分期有关，并在 3郑州大学2002年硕士毕业论文 伪母细胞痈中VE口F和＊OS e与血管生成的关系及临床意义促进肿瘤血管形成中起到重要作用。3、eNOS表达与其组织病理分型及临床分期有关，并在促进肿瘤血管形成中起重要作用。4、VEGF在肾母细胞瘤表达与eNOS表达之间比较无明显相关。5、本研究为肾母细胞瘤的诊断和治疗提供一种新的思路口更多还原

【Abstract】 Wilms’ tumor is one of the most common abdominal malignant tumor in children.In 1899,German doctor Wilms reported this disease for the first time,so it was named after his name.Because it is generated from rear renal embryo in embryogenesis and the tumor are consisted of cells that looking like nephrogenic cellsjt is also called nephroblastoma.In children ,its incidence is only second to that of neuroblastoma in some countries but is a little higher than the laters in china.The tumor usually generate in the first five years,especially at the age of 2 - 4years.lt often generated with congenital malformation,such as congenital lacking of iris,congenital limbs hypertrophy in one side.The tumor generate from indifferential renal embryo and can form every part of nephridium.This mixed malignant tumor is consisted of mesenchymal tissue and epithelial tissue,andnowadays its main prognosis factors are the histological strutcure,age,the totally excision of original tumor, metastasis and two sides of tumor. We still don’t find specific diagnostic marker in nephroblastoma now.By means of immunohistochemical techniques, to investigate the expression of endothelial nitric oxide synthase (eNOS) and vascular endothelial growth factor (VEGF), and to study their relationships in nephroblastoma and the their role in the angiogenesis in nephroblastoma,to reveal the relationships among them and the biogical characters of this tumor . All these will provide people with new thoughts and basis to prevent and treat cancer.Materials and methods: we have collected 50 children’s nephroblastoma specimens from surgical resections, and all sections were formalin-fixed and paraffin embedded. The patients, aged from 4 months to 15 years , average 5.44+4.73 years, 32 males and 18 females, were not performed any radiotherapy and chemotherapy before operation.Their HE staining sections were affirmed and histopathologic types were classified. Their clinical stage were classified according to the condition during operation and the pathologic diagnosis after operation (by NWTS-3 Criteria). Among them ,30 patients had stage I-II , 20 stage III-IV . 40 patients had FH (favorable histology) type tumor, 10 UH (unfavorable histology) type . The expression of MVD, eNOS andVEGF in 50 patients with nephroblastoma were studied retrospectively by using immunohistochemical staining (SABC). The immunohistochemical staining of eNOS, VEGF were assessed according to this criterion: those having positive staining in less than ten percent of tumor cells were graded as "- " ,more than ten percent as "+". MVD was assessed by the mean of blood vessel quantity of five"hot spof’under 400 x microscope. Statistical analysis was executed by SPSS 10.0 software. P values less than 0.05 were considered significant.Results: l.The MVD in 10 patients (UH type) was 85.92 + 14.36 which was significantly higher than that in 40 patients (FH type) was 50.57+ 13.01(PO.05); The MVD in 20 patients (III-IV stage) was 70.39+17.59 which was significantly higher than that in 40 patients ( I-II stage) was 49.14+15.70 (PO.05). 2.The expression rate of VEGF was 80.00% in patients (III-IV stage) which was significantly higher than that in patients ( I-II stage), (P<0.05). The expression rate of VEGF was 36.67% in patients ( I-II stage) .The expression rate of VEGF was 65.00% in FH type while it was 60.00% in UH type (P>0.05).The MVD in VEGF positive group was 65.70+16.88 and was higher than that of VEGF negative group (PO.05). 3.The expression rate of eNOS was 70.00% in patients(UH type) which was significantly higher than that in patients(FH type) (P<0.05) . The expression rate of eNOS was 27.50 % in patients(FH type). The expression rate ofeNOS was 60.00% in III-IV stage and expression rate of eNOS while it was 20.00% in I-II stage, (P<0.05).The MVD in eNOS positive group was 73.13 + 19.38 and was higher than that of eNOS negative group, (P<0.05). 4.There was no relation between the expression of VEGF and the expression of eNOS in nephroblastoma. 5. The expression更多还原