Detection of pigment epithelium derived factor gene mutation in a human malignant melanoma cell line A375
- VernacularTitle:人恶性黑素瘤细胞株A375中色素上皮衍生因子基因的突变
- Author:
Chengfeng ZHANG
;
Leihong XIANG
;
Kun GUO
;
Yinkun LIU
;
Zhizhong ZHENG
- Publication Type:Journal Article
- Keywords:
Melanoma,experimental;
Angiogenesis inhibitors;
Genes,PEDF;
Mutation
- From:
Chinese Journal of Dermatology
2008;41(10):680-682
- CountryChina
- Language:Chinese
-
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 ofpolymerase 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.
