Analysis of mutation in the neurofiblomatosis type 2 (NF2) gene in preneoplastic and neoplastic lesion of human pleura.

(背景与目的)神经纤维瘤病2型(NF2)基因突变在人类恶性恶性淋巴瘤中有较高的发生率。为建立恶性间皮瘤的NF2基因治疗模型，我们对恶性间皮瘤和手术切除的恶性间皮瘤癌旁胸膜进行了NF2基因的基因分析。并对肺癌恶性恶性胸膜炎和恶性间皮瘤进行免疫组织化学分析，以评价NF2基因突变与肿瘤生物学行为的关系。(材料和方法)对3例恶性恶性淋巴瘤和65例肺癌切除后胸膜亚临床播散的病例进行了检查。用酚-氯仿法提取肿瘤DNA，用聚合酶链式反应-单链构象多态性方法检测肿瘤组织中NF2基因突变。NF2基因的C-18和A-19抗体，Ki-67抗原的MIB-1抗体和p53癌蛋白的DO-7抗体进行免疫组织化学染色。(结果与结论)3例NF2基因外显子3、外显子4、外显子7、外显子13和外显子16均成功获得基因组扩增产物。但对这5个外显子进行SSCP分析，未发现NF2基因明显突变。免疫组织化学分析显示C-18和A-19抗体也有相同的结果。2例恶性淋巴瘤Ki-67标记指数低，P53蛋白表达阴性。在肺癌患者中，Ki-67标记指数是独立的预后因素，且与肺癌胸膜播散者的临床病程密切相关。要评估NF2基因在恶性间皮瘤患者中的作用，还需要对更多的恶性间皮瘤病例进行进一步研究。

(Background and aim) It was reported high incidence of mutation in the neurofiblomatosis type 2 (NF2) gene in human malignant methothelioma. To establish a model of gene therapy with NF2 gene for malignant methothelioma, we performed genetic analysis of NF2 gene with DNA of malignant methothelioma and tumor adjacent pleura of surgically resected malignant methothelioma cases. And we also performed immunohistochemical analysis of these malignant methothelioma and malignant pleurisy cases of lung cancer to evaluate mutation of NF2 gene and biological behavior of the tumor. (Materials and Methods) Three cases of resected malignant methothelioma and 65 cases of resected lung cancer cases with subclinical pleural dissemination were examined. Tumor DNA was isolated by phenol-chloroform method and mutation of NF2 gene was evaluated by PCR-SSCP method. Immunohistochemistry was performed with C-18 and A-19 antibody for NF2 gene, MIB-1 antibody for Ki-67 antigen and DO-7 antibody for p53 oncoprotain. (Results and Conclusion) In these three cases, genomic PCR amplification was successfully obtained in the exon 3, exon 4, exon 7, exon 13 and exon 16 of NF2 gene. But no apparent mutation of NF2 gene was detected by SSCP analysis which was performed in these five exons. Immunohistochemical analysis also showed the same results with C-18 and A-19 antibodies. Ki-67 labeling index was low and negative for p53 oncoprotain in two methothelioma cases. In lung cancercases, it was disclosed Ki-67 labeling index was independent prognostic factor and correlated well with clinical course of the lung cancer cases with pleural dissemination. Further investigation with more methothelioma cases must be needed to evaluate the role of NF2 gene in malignant methothelioma patients.