摘要
胰腺癌早期诊断困难,恶性程度高,手术切除率低,预后极差。在美国1997年死于胰腺癌的患者超过28000人,居癌症死亡的第5位。胰腺癌确诊后平均5年生存率低于5%。近年来随着各种治疗技术和诊断方法的发展,肿瘤的诊断率和生存率正在逐年提高,但胰腺癌的预后仍然很差。因此对于胰腺癌分子机制的研究,特别是寻找胰腺癌诊断和治疗的靶点及与胰腺癌调控相关的信号传导通路具有重要的意义。
随着人类基因计划的初步完成,蛋白质组学成为目前国际上的研究热点。与基因相比,蛋白质是人体的各种生命活动的直接执行者。通过对肿瘤组织与正常组织所表达的蛋白质的对比分析,可以找出其中的差异蛋白质。这些差异蛋白质通过其所在的信号传导通路调控肿瘤的发生和发展。
国际上出现的各种先进的实验技术和分析方法大大促进了蛋白质组学的发展。首先是高通量蛋白质筛选技术,如二维电泳、组织芯片、PBSⅡ/C型表面增强激光解吸离子化飞行时间质谱系统、pathway array分析等,这些方法的特点是可以同时检测大量的蛋白质,提高检测效率。高通量蛋白筛选技术能使我们在短时间内获得大量的数据,但传统的统计方法无法分析如此大量的数据。国际上通用的解决办法是将基因分析的技术(如软件和数据库等)用于分析蛋白质的功能和信号传导通路,可以有效的解决这一问题。基因芯片显著性分析(SAM)、基因芯片分析软件包(BRB-ArrayTools)、信号网络分析(Ingenuity Pathway Analysis, IPA)等技术目前常用于高通量蛋白质数据的分析。
使用表皮生长因子受体(EGFR)抑制剂治疗结直肠癌、非小细胞肺癌和乳腺癌的临床效果已经得到公认,目前EGFR抑制剂在胰腺癌治疗中的作用成为国内外研究的热点。西妥昔单抗和埃罗替尼已经被FDA批准用于临床治疗晚期胰腺癌。只有K-ras基因野生型的患者对EGFR抑制剂治疗敏感,因为K-ras基因突变会持续激活EGFR信号传导路的下游,从而使EGFR抑制剂的抑制作用不能向下传递。在这一过程中EGFR通路及其它信号传导通路的蛋白质表达情况国内外未见相关报道。
我们采用pathway array分析方法,筛选出胰腺癌组织和正常胰腺组织之间的差异蛋白,通过SAM、BRB-ArrayTools、IPA等统计分析软件,确定了12个有统计学意义的差异蛋白及其主要涉及的信号传导通路。同时测定了胰腺癌标本的K-ras基因突变情况,并对其对蛋白质表达的影响进行了分析。我们也结合标本的临床资料,探讨了胰腺癌患者蛋白质表达与年龄、性别、种族和淋巴结转移情况之间的关系。
Purpose:
Pancreatic cancer is the fourth leading death of cancer among men and women in the United States The overall survival rate is less than 1% at 5 years with most patients dying within one year[9]. The lack of diagnosis early method and effective treatment is the main problem to pancreatic cancer. We aim to identify protein signatures of pancreatic cancer for diagnosis and therapy.
Materials and Method:
we employed protein pathway array method to assess the protein expression of 11 paires pancreatic cancer samples,5 pancreatic samples and 2 normal pancreatic samples. Next, we use high resolution melting (HRM) curve analysis for KRAS mutation of all pancreatic cancer samples. In addition, Pathways associated with these expression changes were identified using the Ingenuity Pathway Analysis tool.
Results and Conclusion:
8 proteins were upregulated and 4 proteins were downregulated in pancreatic cancer compared with and normal tissues. KRAS mutations were identified in 9 of 15 tumor tissues (60%). p-PKCα/βⅡin K-ras positive upregulation. diffirent express protein mainly affects cell cycle, cellar Growth and Proliferation, Tissue Morphology network functions.IPA result demonstrated the proteins of our study is correlation with lots of signaling pathway.
Eight proteins (FAS, cdk2, cdc2 p34, EGFR, Akt, Vimentin, E-cadherin ICAM-1) were up-regulated and four protein (cyclin B1, Cdc25B, H-Ras, HIF-3α,) were down-regulated in pancreatic cancer compared with normal tissues.these differentially expressed proteins affect cell proliferation, cell cycle and apoptosis ect.these proteins can be used to early diagnosis and therapeutic target.
引文
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