McKie A B, Iwamura T, Leung H Y, Hollingsworth M A, Lemoine N R
Imperial Cancer Research Fund Oncology Unit, Royal Postgraduate Medical School, Hammersmith Hospital, London, United Kingdom.
Genes Chromosomes Cancer. 1997 Jan;18(1):30-41. doi: 10.1002/(sici)1098-2264(199701)18:1<30::aid-gcc4>3.0.co;2-2.
DNA fingerprinting can be used to detect genetic rearrangements in cancer that may be associated with activation of oncogenes and inactivation of tumour suppressor genes. We have developed a fingerprinting strategy based on polymerase chain reaction (PCR) amplification of genomic DNA with primers specific for the Alu repeat sequences, which are highly abundant in the human genome. This has been applied to DNA from pancreatic cancer and paired normal samples to isolate and identify fragments of genomic DNA rearranged in the malignant cells. These fragments have been sequenced and used as probes to isolate hybridising clones from gridded bacteriophage P1, phage artificial chromosome, and cosmid libraries for fluorescent in situ hybridisation mapping and the identification of expressed sequences. Further characterisation has identified a putative novel gene (ART1) that is up-regulated specifically in pancreatic cancer as well as another sequence with similarity to genes involved in differentiation (POU domains). In conclusion, we suggest that Alu-PCR fingerprinting may be a useful technique for the identification of genes involved in tumourigenesis.
DNA指纹图谱可用于检测癌症中的基因重排,这些重排可能与癌基因的激活和肿瘤抑制基因的失活有关。我们基于聚合酶链反应(PCR)开发了一种指纹图谱策略,该反应使用针对Alu重复序列的特异性引物对基因组DNA进行扩增,Alu重复序列在人类基因组中高度丰富。这已应用于胰腺癌DNA和配对的正常样本,以分离和鉴定恶性细胞中重排的基因组DNA片段。这些片段已被测序,并用作探针从网格化的噬菌体P1、噬菌体人工染色体和黏粒文库中分离杂交克隆,用于荧光原位杂交作图和鉴定表达序列。进一步的表征鉴定出一个推定的新基因(ART1),该基因在胰腺癌中特异性上调,以及另一个与参与分化的基因(POU结构域)相似的序列。总之,我们认为Alu-PCR指纹图谱可能是一种用于鉴定参与肿瘤发生的基因的有用技术。