【作者】 景国慧；

【导师】 王燕；

【作者基本信息】 天津医科大学 ， 内科学， 2010， 硕士

【摘要】 目的：通过检测wwox基因6-8外显子mRNA在良、恶性胸腔积液中的表达,探讨WWOX基因在良、恶性胸腔积液临床鉴别诊断中的价值。方法：根据病理学诊断结果,将来自2007年9月-2008年6月天津胸科医院胸内科住院患者的76例胸腔积液标本分为两组,其中良性胸腔积液组20例,肺癌合并恶性胸腔积液组56例。胸穿抽取胸腔积液10ml,提取RNA；经逆转录合成cDNA,再经PCR反应,用电泳直接检测cDNA的扩增产物用逆转录-聚合酶链反应(RT-PCR)技术进行良、恶性性胸腔积液WWOX基因mRNA6-8外显子表达的检测。进一步分析恶性胸腔积液患者WWOX6-8外显子表达与临床特征(包括性别、吸烟情况、肿瘤组织类型、分期)的关系。数据用SPSS13.0软件处理,两样本率的比较用四格表资料的确切概率法(Fisher’s精确概率法)及χ2检验法,P<0.05为有显著性差异。结果：1.恶性胸腔积液组：56例标本中,39例(69.6%)标本未能扩增出WWOX基因6-8外显子片段；而56例标本中,其中31例经胸水细胞学及胸水沉淀物病理确诊肺癌的样本均未能扩增出WWOX基因6-8外显子片段,而胸水细胞学及胸水沉淀物病理阴性,最后经胸膜活检证实肺癌的25例恶性胸水样本中,有8例未能扩增出WWOX基因6-8外显子片段。良性胸腔积液组：20例标本全部(100%)扩增出WWOX基因6-8外显子片段。经统计方法处理,χ2值为6.765(p<0.05),两者相比较差异有统计学意义。胸腔积液WWOX基因6-8外显子转录本缺失诊断恶性胸腔积液的敏感度为69.6%,特异度为100%,阳性预测值为100%,阴性预测值为100%。2.肺癌并恶性胸腔积液WWOX基因6-8外显子缺失率在性别、是否重度吸烟及肿瘤组织类型(非小细胞肺癌与小细胞肺癌)有显著性差异(分别p=0.001、p=0.001、p=0.041),但与肿瘤临床分期无显著性差异(p=0.27)。男性、长期大量吸烟者及非小细胞肺癌肺癌患者恶性胸腔积液WWOX基因6-8外显子缺失率较高。结论：研究表明,位于16q23.3-24.1的WWOX基因在恶性胸腔积液中存在较高的6-8外显子转录本缺失率,提示WWOX基因可能在恶性胸腔积液的发生、发展中起一定作用；WWOX基因6-8外显子转录本的丢失是恶性胸腔积液的分子标志之一；WWOX基因6-8外显子检测可作为鉴别良、恶性胸腔积液辅助手段更多还原

【Abstract】 Objective:To detect the abnormal expression of WWOX (WW domain containing oxidoreductase) exons 6-8 In human lung benign and malignant pleural effusion at mRNA leval and to draw some conclusion concering whether loss of WWOX exons 6-8 play an role in the tumorigenesie of malignant pleural effusion.Methods:76 pleural effusion samples come from Tianjin Chest Hospital Medical Department in 2007 September to 2008 June in, According to cytology results samples were divided into two groups:20 in benign effusions and 56 in malignant effusions which from lung cancer. Take 10ml of pleural effusion, extracted total RNA, react cDNA by RT, after PCR 5μl of sample electrophoresis, observe and photographed in ultraviolet light. To analysis the relationship of the expression of WWOX exons 6-8 with clinical charactors (gender, smoking, histological type, stage),we use chisquare test and exact probability test of contingency table by spss 13.0 sofeware, P<0.05 have statistics difference.Results:(1) malignant effusions groups,39 (69.6%) specimens failed PCR amplification of DNA fragments of the expected length,in 56 cases of malignant pleural effusion,39 specimens failed to amplify exon 6-8 WWOX gene fragment, while the samples of 20 (100%) patients with benign pleural effusions WWOX gene was amplified exon 6-8 fragment (χ2 is 6.765,p<0.05).31 cases of pleural effusion by pleural fluid cytology and pathological diagnosis of lung cancer sediment samples cannot amplificate exon 6-8 WWOX gene fragments, and pleural fluid cytology and pleural effusion sediment pathology negative.Finally, in 25 cases of malignant pleural effusion of lung cancer samples confirmed by pleural biopsy,8 cases failed to amplify exon 6-8 WWOX gene fragments.(2) We further observed the transcript loss of WWOX exons 6-8 was significantly higher in the men compared to the women (p=0.001), it is also higher in the smokers compared to the none-smokers (p=0.001)and squamous cell carcinomas compare to adenocarcinomas(p=0.041), But it was not associated with TNM stage(p=0.27). There is higher transcript loss of WWOX exons 6-8 in male,heavily smoking,non small cell lung cancer patients.Conclusion:The study shows that, at 16q23.3-24.1 of the WWOX gene in non-small cell lung cancer there is a higher transcript exon 6-8 deletion rate,it suggests that WWOX gene may play an important role in the development of malignant pleural effusion,the loss of WWOX gene transcripts exon 6-8 is an molecular marker of malignant pleural effusion; WWOX gene exon 6-8 mRNA can be used as indicators in identifying benign and malignant pleural effusion.更多还原