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目的 寻找与B16黑色素瘤发生相关的DNA序列或片段。方法 用 10 5条引物对B16黑色素瘤及其来源小鼠C57BL/ 6J基因组DNA进行随机扩增多态DNA分析 ,比较肿瘤组织及其相应正常组织的DNA指纹 ,对差异很明显的片段回收、克隆和测序 ,序列与GenBank数据库进行同源性分析。结果 10 5条引物中有 2 4条引物扩增出的条带在肿瘤组织与其相应正常组织间存在差异。在 6个差异很明显的回收DNA片段中 ,引物AB8 5扩增后所得差异片段B8 5 ,肿瘤组织中此片段缺失 ,该片段序列长 610bp ,与GenBank序列数据库中鼠肌色素上皮源性因子基因具有 99% ( 4 19/ 4 2 1)的同源性 ,与鼠肌色素上皮源性因子 (PEDF)mRNA的同源性为 10 0 % ( 2 13 / 2 13 ) ,可以认为该序列即为此基因。结论 黑色素瘤中存在PEDF基因缺失 ,提示PEDF基因与黑色素瘤发生相关。
Objective To search for DNA sequences or fragments associated with the development of B16 melanoma. Methods A random amplified polymorphic DNA analysis of B16 melanoma and mouse C57BL/6J genomic DNA was performed using 105 primers. The DNA fingerprints of tumor tissues and their corresponding normal tissues were compared, and the fragments with obvious differences were recovered and cloned. And sequencing, sequence homology analysis with GenBank database. RESULTS: Among the 10 primers, 24 bands were amplified by 24 primers and there were differences between the tumor tissues and their corresponding normal tissues. In 6 remarkably recovered DNA fragments, the differential fragment B8 5 amplified by primer AB8 5 was deleted from the tumor tissue. This fragment was 610 bp in length and was associated with mouse myoepithelial-derived factor genes in GenBank sequence database. With 99% (4 19 / 4 21) homology, the homology with murine myoepithelial-derived factor (PEDF) mRNA is 10% (2 13 / 2 13). This sequence can be considered as This gene. Conclusion There is a deletion of PEDF gene in melanoma, suggesting that PEDF gene is associated with melanoma.