Yasuda J, Kashiwabara H, Kawakami K, Uematsu K, Sugano K, Perucho M, Sekiya T
Oncogene Division, National Cancer Center Research Institute, Tokyo, Japan.
Biol Chem. 1996 Sep;377(9):563-70. doi: 10.1515/bchm3.1996.377.9.563.
The microsatellite mutator phenotype (MMP), detected as a change in the number of repeating units in hundreds of thousands of microsatellite sequences in the tumor cell genome, underlies the carcinogenesis of a variety of tumors including sporadic and hereditary nonpolyposis colon cancers. This enhanced microsatellite instability was discovered using arbitrarily primed polymerase chain reaction (AP-PCR) fingerprinting of DNA from colon cancers. In this study, we found an arbitrary primer that can amplify multiple DNA fragments containing repeated sequences, including the poly A tracts found in the Alu repeats of the human genome. The combined use of primer labeling with fluorescence and an automated DNA sequencing analysis of AP-PCR products (FAP-PCR) detected alterations in fingerprint bands in all DNA samples previously determined to belong to the MMP. Fluorescent AP-PCR fingerprinting using this single arbitrary primer provides a convenient and efficient method for detecting tumor specific fingerprint alterations that are usually undetectable by conventional fingerprinting.
微卫星突变体表型（MMP）表现为肿瘤细胞基因组中数十万个微卫星序列重复单元数量的变化，是包括散发性和遗传性非息肉病性结肠癌在内的多种肿瘤发生的基础。这种增强的微卫星不稳定性是通过对结肠癌DNA进行任意引物聚合酶链反应（AP-PCR）指纹分析发现的。在本研究中，我们发现了一种任意引物，它可以扩增多个包含重复序列的DNA片段，包括人类基因组Alu重复序列中的多聚A序列。引物荧光标记与AP-PCR产物的自动DNA测序分析（FAP-PCR）相结合，在先前确定属于MMP的所有DNA样本中检测到指纹条带的改变。使用这种单一任意引物的荧光AP-PCR指纹分析为检测肿瘤特异性指纹改变提供了一种方便、高效的方法，而这些改变通常是传统指纹分析无法检测到的。
