Nonradioactive arbitrarily primed polymerase chain reaction: a novel technique for detecting genetic defects in skin tumors.
Nonradioactive arbitrarily primed polymerase chain reaction: a novel technique for detecting genetic defects in skin tumors.
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非放射性任意引发聚合酶链反应：一种检测皮肤肿瘤遗传缺陷的新技术。
DOI：
10.1111/1523-1747.ep12329949
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Schlegel,J
中科院分区：
文献类型：
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作者：
Vogt,T;Stolz,W;Landthaler,M;Rüschoff,J;Schlegel,J
黑素细胞性和非黑素细胞性皮肤肿瘤的发生和发展伴随着多种遗传缺陷，并可能由多种遗传缺陷引起。然而，与其他人类肿瘤相比，皮肤癌中有限的可用组织通常会阻碍对早期病变的天然物质进行广泛的遗传研究。因此，我们采用了一种新的DNA指纹技术，基于聚合酶链反应（PCR），任意引物PCR（AP-PCR）。这项技术使我们能够扫描基因组的大部分（约30 kb/PCR反应），以从微量组织开始进行体细胞突变。与以前的报告AP-PCR，我们能够可视化PCR产物的快速非放射性银染色技术，使用一个简单的设备染色的大型聚丙烯酰胺凝胶。在9个良性和恶性黑色素细胞皮肤肿瘤中，该方法提供了一组可重复的DNA指纹。通过比较肿瘤细胞和血液白细胞组成型DNA的指纹图来检测遗传缺陷。由于非放射性AP-PCR指纹也提供了独特的能力，分离和序列多态性DNA片段的指纹凝胶，我们得出结论，这种技术似乎是重要的，实际上是可行的，阐明皮肤肿瘤的遗传根源。
Initiation and progression of melanocytic and nonmelanocytic skin tumors are accompanied and probably caused by a variety of genetic defects. In contrast to other human tumors, however, limited amounts of available tissue in skin cancer often hamper extensive genetic studies of native material of early lesions. Therefore, we applied a novel DNA fingerprinting technique based on polymerase chain reaction (PCR), the arbitrarily primed PCR (AP-PCR). This technique enabled us to scan large parts of the genome (about 30 kb/PCR reaction) for somatic mutations starting with minute amounts of tissue. In contrast to previous reports on AP-PCR, we were able to visualize PCR products by a rapid nonradioactive silver-staining technique using a simple device for staining of large polyacrylamide gels. In nine benign and malignant melanocytic skin tumors, the method provided a set of reproducible DNA fingerprints. Genetic defects were detected by comparing the fingerprints of tumor cells and constitutive DNA from blood leukocytes. Since nonradio-active AP-PCR fingerprinting also offers the unique capability to isolate and sequence polymorphic DNA fragments from fingerprint gels, we conclude that this technique seems to be important and practically feasible for elucidating the genetic roots of skin tumors.