Epigenetic silencing of the PTEN gene in melanoma.
Phosphatase and tensin homologue deleted from chromosome 10 (PTEN) seems to be an important tumor suppressor gene in melanoma. Because the PTEN gene is only infrequently deleted or mutated, and because the PTEN protein is low to absent in a significant number of melanomas, we investigated alternative methods of epigenetic silencing. We did quantitative positional methylation analysis (pyrosequencing) on 37 sera from melanoma patients and on 21 pairs of corresponding sera and melanoma specimens in addition to Taqman reverse transcription-PCR. We report significant positional PTEN promoter methylation in 62% of circulating DNA isolated from sera of patients with metastatic melanoma. The percentage of methylation of a selected CpG island in blood showed a correlation with methylation levels in the corresponding melanoma tissue. Moreover, high percentages of PTEN methylation were associated with low PTEN transcription levels. Using the demethylation agent 5-aza-2'-deoxycytidine, reduced methylation and a corresponding increase in PTEN protein were observed in BLM melanoma cells, leading to reduced AKT activity in an in vitro kinase assay. In summary, epigenetic PTEN silencing seems to be a relevant mechanism of inactivating this tumor suppressor gene in melanoma that may promote melanoma development by derepression of the AKT pathway.
Mirmohammadsadegh
Alireza
A
Department of Dermatology, Heinrich-Heine University, Moorenstrasse 5, D-40225 Düsseldorf, Germany.
Marini
Alessandra
A
Nambiar
Sandeep
S
Hassan
Mohamed
M
Tannapfel
Andrea
A
Ruzicka
Thomas
T
Hengge
Ulrich R
UR
D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D002879:Chromosomes, Human, Pair 10 / Q000235:genetics; D018899:CpG Islands / Q000235:genetics; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000097:blood / Q000235:genetics; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D016147:Genes, Tumor Suppressor; D006801:Humans; D008297:Male; D008545:Melanoma / Q000097:blood / Q000235:genetics* / Q000473:pathology / Q000556:secondary; D008875:Middle Aged; D008969:Molecular Sequence Data; D051059:PTEN Phosphohydrolase / Q000096:biosynthesis / Q000235:genetics*; D011401:Promoter Regions, Genetic
D004273:DNA, Neoplasm; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human