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Mutation analysis of tumors in ophthalmological area to to choose the treatment modalities---mutation analysis using paraffin specimens---

眼科肿瘤突变分析以选择治疗方式---石蜡标本突变分析---

基本信息

批准号：
13671819
负责人：
YAMASHITA Hidetoshi
金额：
$ 2.24万
依托单位：
Department of Ophthalmology and Visual Science, Yamagata University School of Medicine
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

Purpose : To evaluate the usefulness of gene mutation analysis and genotype analysis to diagnose tumors in ophthalmological area. The target genes are TGF-β and immunoglobulin light chain.Methods : [mutation analysis] Expression and mutations of TGF-β type II receptor (T β R2), Smad 4 (one of the signal transducers) were investigated. DNA was extracted from paraffin embedded specimens from 21 patients, which included malignant melanoma, sebaceous carcinoma, squamous cell carcinoma, basal cell carcinoma, pleomorphic adenocarcinoma, pleomorphic adenoma, solitary fibrous tumor, hemangioma and others. DNA fragments which encode some exons of TβR2 and Smad4 were amplified by polymerase chain reaction (PCR), and then single strand conformation polymorphism analysis (SSCP) was performed to detect mutations. [Expression of TβR2 and Smads] Expression was observed immunohistochemically in the paraffin sections. [immunoglobulin rearrangement] Three cases of lymphoproliferative lesions were treated at Yamagata University Hospital. The surgical biopsy specimens were examined histopathologically and Southern blot hybridization was used to detect immunoglobulin rearrangement as a genotype analysis.Results : Two out of 21 cases showed the band shift of TβR2 exon 6 and smad 4 exon 11a by SSCP. The expression of TβR2 was shown to decrease by immunohistochemically. In one case of lymphoproliferative tumors, rearrangement of immunoglobulin light chain gene. This case was shown to be malignant.Conclusions : This study suggests that the mutation in TGF-β related genes may be related to the tumorigenesis, however, may not be used to diagnose malignancy clinically. The rearrangement of immunoglobulin light chain gene is very useful to diagnose malignancy.

目的：评估基因突变分析和基因型分析在眼科肿瘤诊断中的有用性。靶基因为TGF-β和免疫球蛋白轻链。方法：[突变分析]研究TGF-βII型受体(TβR2)、Smad 4(信号转导蛋白之一)的表达和突变。从21例患者的石蜡包埋标本中提取DNA，包括恶性黑色素瘤、皮脂腺癌、鳞状细胞癌、基底细胞癌、多形性腺癌、多形性腺瘤、孤立性纤维瘤、血管瘤等。通过聚合酶链式反应（PCR）扩增编码TβR2和Smad4部分外显子的DNA片段，然后进行单链构象多态性分析（SSCP）检测突变。 [TβR2和Smads的表达] 通过免疫组织化学法观察石蜡切片中的表达。【免疫球蛋白重排】山形大学医院收治3例淋巴增生性病变。对手术活检标本进行组织病理学检查，并采用Southern印迹杂交检测免疫球蛋白重排作为基因型分析。结果：21例中2例经SSCP检测显示TβR2外显子6和smad 4外显子11a带移动。免疫组织化学显示TβR2的表达降低。在淋巴增殖性肿瘤的一例中，免疫球蛋白轻链基因重排。结论：本研究提示TGF-β相关基因的突变可能与肿瘤的发生有关，但不能用于临床诊断恶性肿瘤。免疫球蛋白轻链基因的重排对于诊断恶性肿瘤非常有用。