ISOLATION AND CHARACTERIZATION OF ALLELIC LOSSES AND GAINS IN COLORECTAL TUMORS BY ARBITRARILY PRIMED POLYMERASE CHAIN-REACTION
ISOLATION AND CHARACTERIZATION OF ALLELIC LOSSES AND GAINS IN COLORECTAL TUMORS BY ARBITRARILY PRIMED POLYMERASE CHAIN-REACTION
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DOI：
10.1073/pnas.89.21.10065
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发表时间：
1992-11-01
影响因子：
11.1
通讯作者：
PERUCHO, M
中科院分区：
文献类型：
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作者：
PEINADO, MA;MALKHOSYAN, S;PERUCHO, M
任意引物聚合酶链反应（AP-PCR）[Welsh，J. & McClelland，M.（1990）Nucleic Acids Res. 18，7213-7218]已用于检测结肠和直肠肿瘤中的体细胞遗传改变。比较了同一个体的正常组织和肿瘤组织的单引物DNA指纹图谱。在用相同的任意引物重新扩增后，已经克隆了显示肿瘤组织DNA中相对于正常的降低和增加的强度的AP-PCR条带。标准限制性片段长度多态性和Southern印迹分析表明，这些DNA序列分别在肿瘤细胞基因组中经历了等位基因丢失和获得。通过仓鼠体细胞/人细胞杂交的PCR和连锁分析，将缺失的序列定位于17号染色体的短臂。这些结果显示了AP-PCR在单个步骤中检测和分离代表癌细胞非整倍性基础的两种遗传改变的DNA序列的能力：杂合性丢失和染色体获得。总之，他们还显示了定量性质的扩增水平在体外获得的AP-PCR，从而提供了一个替代的分子方法的基础上，癌症细胞遗传学。
The arbitrarily primed polymerase chain reaction (AP-PCR) [Welsh, J. & McClelland, M. (1990) Nucleic Acids Res. 18, 7213-7218] has been used to detect somatic genetic alterations in tumors of the colon and rectum. DNA fingerprints generated by single arbitrary primers were compared between normal and tumor tissue of the same individuals. AP-PCR bands showing decreased and increased intensities in tumor tissue DNA, relative to normal, have been cloned after reamplification with the same arbitrary primer. Standard restriction fragment length polymorphism and Southern blot analyses show that these DNA sequences have undergone allelic losses and gains, respectively, in the tumor cell genome. The deleted sequences have been assigned to the short arm of chromosome 17 by PCR of somatic hamster/human cell hybrids and linkage analysis. These results show the ability of the AP-PCR to detect and isolate, in a single step, DNA sequences representing two of the genetic alterations that underlie the aneuploidy of cancer cells: losses of heterozygosity and chromosomal gains. Altogether, they also show the quantitative nature of the amplification levels obtained in vitro by AP-PCR, which thus provides the basis for an alternative molecular approach to cancer cytogenetics.