【作者】 王光辉；

【导师】 杜贾军；

【作者基本信息】 山东大学 ， 外科学， 2010， 硕士

【摘要】 目的肿瘤的侵袭转移是肿瘤患者死亡的原因之一,因此抑制肿瘤的侵袭转移是治疗肿瘤最重要的策略之一。研究表明,Twist蛋白是属于碱性的螺旋-环-螺旋蛋白家族中的高度保守的转录因子,可以通过下调肺癌中上皮钙粘素的表达而促进上皮间质化。本研究旨在检测Twist和上皮钙粘素在非小细胞肺癌原发灶及转移灶中表达的差异性,以及Twist和上皮钙粘素二者之间的关系,探究Twist和上皮钙粘素在NSCLC发展中的作用。方法1.收集山东大学附属省立医院2000年至2009年11例原发灶和转移灶均被手术切除的非小细胞肺癌病人的石蜡标本和临床资料进行研究。2.采用兔抗人Twist蛋白多克隆抗体和鼠抗人上皮钙粘素单克隆抗体及链霉菌抗生物素蛋白过氧化物酶法检测11例非小细胞肺癌原发灶和11例相关转移灶中Twist和上皮钙粘素的表达。3.结果判定采用半定量积分法：对于Twist,着色程度分为四级：0(阴性),1(弱),2(中等),3(强)。对于上皮钙粘素,着色程度分为五级：0(阴性),1(极弱),2(弱),3(中等),4(强)；据着色程度与阳性细胞数占肿瘤细胞数的百分比的乘积判定阳性等级：5-100(1),101-200(2),201-300(3),和301-400(4)。4.所有数据统计分析均使用统计软件包SPSS17.0处理,Twist和上皮钙粘素二者之间表达的关系采用Spearman检验,原发与转移之间差异性比较采用Wilioxon检验,P<0.05有统计学意义。结果Twist在肺癌组织中高表达,并且与上皮钙粘素表达呈负相关关系。2例(18.2%)病人的原发和转移癌中,Twist表达有差异,其中1例病人在原发癌中的表达强于转移灶。9例(81.8%)病人的原发和转移癌中,上皮钙粘素表达有差异,均为原发灶中的表达强于转移灶。结论上皮钙粘素可以在转移灶中重新表达,且强度明显低于原发灶,Twist在NSCLC转移过程中具有重要作用,其在NSCLC的原发灶及转移灶中均有高表达。Twist可以通过抑制上皮钙粘素表达诱导EMT而促进肿瘤转移。更多还原

【Abstract】 Purpose:Development of metastasis is one of the main causes of lung cancer-related death. It has been known previously that up-regulation of Twist,a highly conserved basic helix-loop-helix transcription factor, in lung cancer cells can promote epithelial to mesenchymal transition through down-regulation of E-cadherin. The aim of this study was to determine if the Twist and E-cadherin status of primary and metastatic lung carcinoma differs and to correlate Twist and E-cadherin expression in primary and metastatic lung cancer specimens.Method:Eleven patients with lung cancer and distant metastases were retrospectively studied,of whom both the primary and metastasis lesions were resected in Provincial Hospital affiliated to Shandong University,Shandong,China, from 2000 to 2009.Immunohistochemistry Streptavidin-Peroxidase(SP)staining was used to detect the expression of Twist and E-cadherin.The sections were scored for twist expression using a four-titer scale: 0, negative;1,weak signal;2,intermediate signal;and 3, strong signal. For E-cadherin,staining intensity was scored as 0(negative),1(trace), 2 (weak),3(intermediate) and 4(strong).Extent of staining was scored according to the percentage of tumor cells with membranous E-cadherin expression in relation to the whole cancer areas(0% to 100%).Then, the scores were multiplied and divided into 4 groups (5 to 100,101 to 200, 201 to 300, and 301 to 400) as corresponding staining degrees(1,2,3, and 4).Correlation analysis between Twist and E-cadherin expression was carried out by comparing total scores for each protein expression in the same sample using Spearman rank test.Wilioxon test was used to compare the Twist and E-cadherin status between primary tumours and related metastatic sites.Result:we demonstrated an overexpression of Twist protein in lung cancer tissues and found that increased Twist expression was correlated with decreased membranous E-cadherin expression. Discordance of Twist expression was observed in two(18.2%) patients. In one case, Twist was expressed higher in the primary tumor than in the metastasis. For E-caherin,nine cases(81.8%)showed primary tumor/metastasis discordance:in nine cases,E-caherin was expressed higher in the primary tumor than in the metastasis.Conclusion:E-cadherin can re-express in metastasis and Twist expression is not stable during metastatic progression. Our data imply that Twist induces EMT on NSCLC by reducing E-caherin expression,thus promoting metastasis.更多还原