中华皮肤科杂志 ›› 2008, Vol. 41 ›› Issue (10): 680-682.

• 论著 • 上一篇    下一篇

人恶性黑素瘤细胞株A375中色素上皮衍生因子基因的突变

张成锋 项蕾红 郭坤 刘银坤 郑志忠   

  1. 复旦大学附属华山医院 上海复旦大学附属华山医院皮肤科 上海复旦大学附属华山医院皮肤科
  • 收稿日期:2008-04-14 修回日期:2008-06-10 发布日期:2008-10-15
  • 通讯作者: 张成锋 E-mail:e3dangdang@hotmail.com

Detection of pigment epithelium derived factor gene mutation in a human malignant melanoma cell line A375

  

  • Received:2008-04-14 Revised:2008-06-10 Published:2008-10-15

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摘要： 目的 探讨色素上皮衍生因子（PEDF）基因在恶性黑素瘤细胞系中的突变情况。方法 采用聚合酶链反应--单链构象多态性分析（PCR-SSCP）对人恶性黑素瘤细胞株A375和正常人黑素细胞中PEDF基因的所有8个外显子进行突变检测。对SSCP分析有异常泳动条带样本的PCR产物进行DNA 测序。结果 人恶性黑素瘤细胞株A375的第3到第7外显子都出现DNA泳动变位，其中以外显子5，6最为明显。PEDF基因第5，6外显子均存在突变。第5外显子的突变类型以单个碱基的缺失为主，第6外显子的突变类型则以单个碱基的置换为主。结论 PEDF基因的突变可能在恶性黑素瘤的发病中起一定作用。

关键词: 黑素瘤, 色素上皮衍生因子, 基因突变

Abstract: Objective To detect the mutations of pigment epithelium derived factor （PEDF） gene in a human malignant melanoma cell line A375. Methods A375 cells and control melanocytes obtained from circumcised prepuce were cultured, genomic DNA was extracted from these cells. All eight exons of PEDF gene were scanned by single strand conformation polymorphism analysis of polymerase chain reaction products （PCR-SSCP） in both A375 cells and control melanocytes. DNA sequencing was performed for the PCR products separated into electrophoretic bands with altered mobility. Results Altered mobility was observed with SSCP analysis in amplicons of exon 3, 4, 5, 6 and 7, with the most obvious alteration occurred in exon 5 and 6. DNA sequencing revealed mutations in both exon 5 and 6. The common type of mutations was single base-deletion in exon 5 and single base-substitution in exon 6. Conclusion Mutations of PEDF gene may contribute to the development of human malignant melanoma.

Key words: Malignant melanoma, Pigment epithelium derived factor