摘要
[背景与目的]结直肠癌是临床常见恶性肿瘤之一,根治性切除术后的患者有近半数会复发和转移,而复发和转移是导致患者预后不良和死亡的最主要原因,结直肠癌患者最常见的远处转移器官是肝脏。基因组DNA的非随机改变在结直肠癌的发生发展中起重要的作用。本研究通过寡核苷酸array CGH分析结直肠腺癌新鲜组织标本,寻找结直肠癌肝转移特异的DNA改变,筛选出可能与肝转移预后密切相关的DNA拷贝数改变和相应的基因。
[材料与方法]收集2006年6月至2006年12月结直肠癌患者根治性切除肿瘤组织标本16例,提取基因组DNA,通过array CGH分析,鉴定结直肠癌的基因拷贝数改变。收集2006年-2007年间根治性切除的结直肠癌120例,构建组织芯片应用免疫组织化学(IHC)方法检测cortactin、HSP70、Ki67、Mcm7、p53、β-catenin、PTP-1βDNAJB6蛋白的表达情况,分析其与肝转移预后的关系。
[结果]Array CGI1结果显示,在结直肠癌中存在7p、8q、13q、19p、20q的增益和4p、4q、8p、17p、18q、21q的缺失。其中13q12.2-q12.3.13q14.11和13q31.3发生高水平扩增；3q26.1、11qll、18q21.2和22q13.2存在纯合性缺失。通过比较基因组改变的频率分布图,我们发现7p、8q和18q在两组间存在最显著频率差异。IHC结果提示：在继发肝转移的结直肠癌中Ki67、MCM7和DNAJB6的阳性表达率分别为46.3%、74.1%和66.7%,均与肝转移相关(Ki67:x2=7.424, P=0.006; MCM7:x2= 24.371, P=0.001; DNAJB6:x2= 4.673, P=0.031)。
[结论]结直肠癌继发肝转移基因拷贝数改变频率高、位点多,Ki67、MCM7和DNAJB6可能作为结直肠癌肝转移的分子标志。
【Background and Purpose】Colorectal cancer is one of the clinical common malignant tumors. Nearly half of the patients who have radical resection will relapse and metastasis, and recurrence and metastasis are the most important causes of poor prognosis and death. The liver is the most common distant metastasis organ of colorectal cancer patients. DNA copy number alterations play an important role in the initiation and progression of colorectal adenocarcinoma. In the present study, we used oligonucleotide array CGH to identify genetic alterations of colorectal cancer with liver metastasis.
[Patients and Methods] The tissue samples were obtained from 16 colorectal cancer tissues. Genomic DNA was isolated, and array CGH was performed with the Agilent Human Genome CGH 8*60K array. Tissue microarray was constructed, containing 120 colorectal cancer tissues. The expression of cortactin. HSP70. Ki67. Mem7. p53.β-catenin. PTP-1B and DNA1B6 proteins were detected by using immunohistochemical method, and the relationships between the proteins expression and liver metastasis of colorectal cancer were analyzed.
[Results] Array CGH showed that the most common genetic changes in colorectal cancer were gains of 7p,8q,13q, 19p,20q and losses of 4p.4q,8p,17p,18q,21q. High level amplifications at 13ql2.2-ql2.3.13q14.11 and 13q31.3 and homozygous deletions at 3q26.1,11q11,18q21.2,22q13.2 were also detected. By comparing the frequency of genomic changes in metastasis and non-metastasis patients.7p,8q and 18q showed most different frequency.. IHC analysis showed that Ki67, MCM7 and DNAJB6 expressed in 46.3%,74.1%and 66.7%of tumors, respectively. The positive expression of Ki67. MCM 7 and DNAJB6 was associated with liver metastases (Ki67:x2=7.424, P=0.006, MCM7: X2=24.371, P=0.001, DNAJB6:%2=4.673, P=0.031).
[Conclusions] There existed lots of chromosomal loci with frequent alterations of DNA copy number in colorectal cancer with liver metastasis. Ki67. MCM7 and DNAJB6 might be useful biomarkers in more advanced stage of colorectal cancer.
引文
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