【作者】 李红梅；

【导师】 周清华； 马力； 梁军；

【作者基本信息】 天津医科大学 ， 肿瘤学， 2010， 博士

【摘要】 p53基因是与人类恶性肿瘤相关性最高的肿瘤抑制基因,是当前癌基因研究热点之一,有资料认为,p53突变普遍存在于肺癌的整个过程中,早期突变阳性的表达高于中晚期肺癌,p53是一种早期筛选的分子指标,且对高危人群的筛选具有一定的前景。端粒酶是一种能以自身RNA为模板,逆转录合成端粒DNA加于染色体末端的核糖核蛋白酶。近年研究表明,肺癌中的高端粒酶活性与肿瘤细胞的高增殖率、病理分期和恶性程度有一定的相关性。国内外研究表明端粒酶在肺癌的早期诊断、治疗和预后预测中有重要意义。本研究为了从肿瘤分子生物学角度探讨多种标本、多基因、多因素联合检测对肺癌早期诊断、病情预后及肿瘤防治方面的意义,我们选择天津医科大学总医院肺癌研究所和青岛大学医学院附属医院肿瘤中心的80例肺癌伴胸腔积液患者和50例肺部良性病变伴胸腔积液患者作为研究对象,采用聚合酶链反应-单链构象多肽性分析(PCR-SSCP)方法和聚合酶链反应-酶联免疫吸附测定法(PCR-TRAP-ELISA)分别检测其外周血、痰、胸腔积液和纤维支气管镜活检组织中p53基因突变率和端粒酶活性。本研究对p53基因突变检测的结果显示：(1)肺癌伴胸腔积液患者外周血、痰、胸腔积液和纤维支气管镜活组织中p53基因突变率均明显高于肺部良性病变伴胸腔积液患者(P<0.01)；(2)不同病理类型、不同分化程度的肺癌伴胸腔积液患者间四种标本的p53基因突变率差异无统计学意义(P>0.05),而吸烟与非吸烟p53基因突变率差异有统计学意义(P<0.05)。(3)p53基因突变率在四种标本间无统计学意义(P>0.05)。(4)四种标本的p53基因突变率联合细胞学诊断肺癌的敏感性高于相应标本的细胞学诊断敏感性,差异有统计学意义(P<0.05)。(5)四种标本联合的p53基因突变检出率高于单一标本p53基因突变检出率,差异有统计学意义(P<0.05)本研究对端粒酶活性检测的结果显示：(1)肺癌伴胸腔积液患者外周血、痰、胸腔积液和纤维支气管镜活组织中端粒酶活性均明显高于肺部良性病变伴胸腔积液患者(P<0.01)；(2)不同病理类型、不同分化程度的肺癌伴胸腔积液患者间四种标本的端粒酶活性差异无统计学意义(P>0.05),而吸烟与非吸烟端粒酶活性差异有统计学意义(P<0.05)。(3)在端粒酶活性四种标本间差异无统计学意义(P>0.05)。(4)四种标本的端粒酶活性联合细胞学诊断肺癌的敏感性高于相应标本的细胞学诊断敏感性,差异有统计学意义(P<0.05)。(5)四种标本联合的端粒酶活性阳性率高于单一标本端粒酶活性阳性率,差异有统计学意义(P<0.05)。本研究对p53基因突变和端粒酶活性联合检测的结果显示：(1)四种标本联合检测p53基因突变率和端粒酶活性高于一种标本的检出率,差异有统计学意义(P<0.05)。(2)联合检测p53基因突变率和端粒酶活性对肺癌伴胸腔积液患者的诊断敏感性高于单项检测敏感性,差异有统计学意义(P<0.05)结论：联合检测外周血、痰、胸腔积液和纤维支气管镜活检组织p53基因突变和端粒酶活性可提高肺癌早期诊断的敏感性和检出率,对判断疾病的性质及病情预后有一定的指导意义。而且临床标本简便易得,患者依从性好。更多还原

【Abstract】 p53 gene is one of the most relevant to human tumor suppressor gene and also is one of the current hot spot in cancer gene research.It is thought that p53 gene mutation is general in the entire process of lung cancer.Also, the mutation of p53 gene is higher in early lung cancers than in advanced ones.Telomerase is a ribonucleoprotein that synthesizes telomeric DNA onto chromosomal ends using an component as a template. Research has shown that the detection of telomerase activity is important to early diagnosis, treatment and prognosis prediction of lung cancer patients.This study is to investigate the significance of multisample, polygene and multifactor combining detection in early diagnosis, prognosis and prevention of lung cancer from the point of view of the tumor molecular biology. We selected 80 lung cancer patients with pleural effusion and 50 benign lung disease patients with pleural effusion from lung cancer institute of tianjin medical university general hospital and cancer center of the affiliated hospital of medical college qingdao university. We employed the technique of PCR-SSCP and TRAP (telomeric repeat amplification protocal)-PCR-ELISA to detect the mutative rates of p53 gene, and the activity of telomerase in peripheral blood, the sputum, pleural effusion and fiberobronchoscopic biopsy of 80 lung cancer patients with pleural effusion and 50 benign lung disease patients with pleural effusion.This study of mutative rates of p53 gene showed as follows:(1) Mutative rates of p53 gene in peripheral blood, the sputum, pleural effusion and fiberobronchoscopic biopsy of 80 lung cancer patients with pleural effusion were higher than 50 benign lung disease patients with pleural effusion significantly. (P<0.01) (2) There was no significant difference in mutative rates of p53 among the different pathologic types and subtype (P>0.05), while there was significant difference among patients of smoking (P<0.05). (3) There was no significant difference in mutative rates of p53 among the different clinical samples (P>0.05) (4) The sensitivity of combined of the detection of p53 point mutative and cytological examination for the four samples was higher than the sensitivity of pole cytological examination significantly (P<0.05). (5) The mutative rates of p53 was significant high in combined four samples than one sample (P<0.05)The study of telomemse activity showed as follows:(1) Telomemse activity in peripheral blood, the sputum, pleural effusion and fiberobronchoscopic biopsy of 80 lung cancer patients with pleural effusion were higher than 50 benign lung disease patients with pleural effusion significantly. (P<0.01) (2) There was no significant difference in telomemse activity among the different pathologic types and subtype (P>0.05), while there was significant difference among patients of smoking (P<0.05). (3) There was no significant difference in telomemse activity among the different clinical samples (P>0.05). (4) The sensitivity of combined of the detection of telomemse activity and cytological examination for the four samples was higher than the sensitivity of pole cytological examination significantly (P<0.05). (5) The telomemse activity was significant high in combined four samples than one sample (P<0.05)The combined detection of p53 point mutative and telomerase activity also gived us the follow results. (1) There was significant higher positive rate of p53 point mutative and telomerase activity in combined four samples than one sample. (2) The sensitivity of combined detection of p53 point mutative and telomemse activity for the four samples was higher than the sensitivity of single examination significantly.Conclusion:The polygene and multifactor combining detection in peripheral blood, the sputum, pleural effusion and fiberobronchoscopic biopsy of 80 lung cancer patients with pleural effusion can improve sensitivity and detectable rate of early lung cancer diagnosis, and has instructive significance for the judgement of the property and prognosis of the disease. The detection of blood, sputum, pleural effusion and fiberobronchoscopic biopsy are safe and convenient. The compliance of patients is good.更多还原