论文摘要
目的:研究结直肠组织Smad4 mRNA及其蛋白表达水平变化、Smad4基因CpG岛异常甲基化与结直肠癌及其临床病理特征的关联性。探讨Smad4蛋白表达异常与结直肠腺瘤、腺癌发生、发展的关系。方法:所有结直肠癌病例均来自福建省立医院2006.11-2007.05外科手术治疗患者,结直肠癌主要指散发型,排除遗传性非息肉病性大肠癌（HNPCC）及家族性结肠息肉病（FAP）癌变。结直肠腺瘤标本取至福建省立医院消化内镜中心内镜下息肉切除术后标本。详细记录病人性别、年龄及结直肠肿瘤易感因素。根据组织标本取材部位和组织学类型,将实验标本分为四组,分别为:（1）腺癌组:标本取自结直肠腺癌病灶处43例;（2）癌旁组:取结直肠腺癌患者的相应远离癌灶的正常粘膜组织（癌旁组织,>5cm）43例;（3）腺瘤组:取腺瘤病灶处32例;（4）健康人黏膜组:取健康人结直肠黏膜组织12例。全部病例均经手术和病理证实。采用RT-PCR方法及测序、半定量RT-PCR方法检测Smad4 mRNA及其表达水平变化;甲基化特异性PCR（MSP）技术检测基因启动子区甲基化程度;免疫组织化学方法检测Smad4蛋白表达水平。应用χ2检验、t检验和Fisher确切概率法对实验结果进行综合分析。结果:1.RT-PCR及测序结果显示43例结直肠癌组织中有25例（58.14%）Smad4 mRNA检测到阳性表达,其阳性表达率低于相应的癌旁组（38/43,88.37%）、腺瘤组（29/32,90.63%）和健康人黏膜组（12/12,100.00%）,差异均有显著性意义（P<0.05）。2.半定量RT-PCR结果显示43例结直肠癌组织中有25例Smad4 mRNA表达水平（0.63±0.25）,其表达水平低于相应的癌旁组（0.95±0.29）、腺瘤组（1.01±0.37）和健康人黏膜组（1.18±0.33）,差异均有显著性意义（P<0.05）。3.MSP示Smad4基因启动子甲基化阳性率分别为60.52%、27.03%、37.50%和16.77%,结直肠癌组明显高于健康人黏膜组和癌旁组织,差异均有显著性意义（P<0.05）,但与腺瘤组差异无显著性（P﹥0.05）。4.免疫组化实验显示Smad4蛋白表达的阳性率分别为44.19%（19/43）,76.74%（33/43）、87.50%（28/32）、91.67%（11/12）,差异均有显著性意义（P<0.05）,并随着浸润深度加深和淋巴结转移,阳性率也随之下降。结论:Smad4的低表达与结直肠癌的发展、生物学行为和预后有相关性。可作为重要的生物学标志及病期评估的标志之一。
论文目录
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