Toma M, Cimponeriu D, Pompilia A, Stavarachi M, Beluşică L, Radu I, Gavrilă L
Departamentul de Genetică Umană, Institutul de Genetică, Universitatea din Bucureşti, România.
J Med Life. 2008 Oct-Dec;1(4):423-8.
Mutations in adenomatous polyposis coli (APC) gene have not been previously characterized among Romanian patients with colorectal cancer (CRC). We initiate this study to detect the mutations in APC gene in blood and tumor samples collected from 16 patients (10 men and 6 women) and blood samples from 21 first and second degree relatives of the patients. For this the presence of mutations in exons 6, 7, 12, 13, 14 as well as in regions B, L and W of exon 15 was investigated using PCR multiplex. In the same time, we have searched for 5 bp deletions at codon 1061 of APC gene by PAGE and SSCP methods. These methods allowed us to evidence identification of the presence of mutations in samples from 7 individuals. In one patient, was detected a deletion of exon 13th of APC gene both in DNA extracted from blood and tumor samples. Multiple deletions (e.g. in exon 6, 12, and in 15L and 15W regions) in DNA extracted from the tumor sample were detected, but not in DNA probe obtained from blood cells. We can speculate that these mutations are an example of genomic instability accompanying the malignancy. Till now, no mutation affecting 1061 codon of APC gene was identified in the patients investigated in our study.
此前尚未对罗马尼亚结直肠癌（CRC）患者的腺瘤性息肉病（APC）基因中的突变进行过特征描述。我们开展这项研究，以检测从16例患者（10名男性和6名女性）采集的血液和肿瘤样本以及21名患者的一级和二级亲属的血液样本中APC基因的突变情况。为此，我们使用多重聚合酶链反应（PCR）研究了第6、7、12、13、14外显子以及第15外显子的B、L和W区域中是否存在突变。同时，我们通过聚丙烯酰胺凝胶电泳（PAGE）和单链构象多态性（SSCP）方法，搜索APC基因第1061密码子处的5个碱基缺失情况。这些方法使我们能够证明在7个人的样本中鉴定出了突变。在一名患者中，从血液和肿瘤样本中提取的DNA中均检测到APC基因第13外显子的缺失。在从肿瘤样本中提取的DNA中检测到多个缺失（例如在第6、12外显子以及第15外显子的L和W区域），但在从血细胞获得的DNA样本中未检测到。我们可以推测，这些突变是伴随恶性肿瘤出现的基因组不稳定性的一个例子。到目前为止，在我们研究的患者中未发现影响APC基因第1061密码子的突变。
