title: Induction of HLA-G expression in a melanoma cell line OCM-1A following the treatment with 5-aza-2'-deoxycytidine.
abstract: The non-classical HLA class I antigen HLA-G is an immune modulator which inhibits the functions of T cells, NK cells, and the Dendritic cells (DC). As a result, HLA-G expression in malignant cells may provide them with a mechanism to escape the immune surveillance. In melanoma, HLA-G antigen expression has been found in 30% of surgically removed lesions but in less than 1% of established cell lines. One possible mechanism underlying the differential HLA-G expression in vivo and in vitro is that the HLA-G gene is epigenetically repressed in melanoma cells in vitro. To test this hypothesis, we treated the HLA-G negative melanoma cell line OCM-1A with the DNA methyltransferase inhibitor 5-aza-2'-deoxycytidine (5-AC) and analyzed whether HLA-G expression can be restored. Our data strongly suggest that HLA-G is silenced as a result of CpG hypermethylation within a 5' regulatory region encompassing 220 bp upstream of the start codon. After treatment, HLA-G mRNA expression was dramatically increased. Western blot and flow cytometry showed that HLA-G protein was induced. Interestingly, HLA-G cell surface expression on the 5-AC treated OCM-1A cells is much less than that on the HLA-G positive JEG-3 cells while a similar amount of total HLA-G was observed. Possible mechanisms for the difference were analyzed in the study such as cell cold-treatment, peptide loading and antigen processing machinery components (APM) as well as beta2 microglobulin (beta2-m) expression. Data revealed that the APM component calreticulin might be involved in the lower HLA-G surface expression on OCM-1A cells. Taken together, our results indicated that DNA methylation is an important epigenetic mechanism by which HLA-G antigen expression is modulated in melanoma cells in vitro. Furthermore, to the first time, we hypothesized that the deficiency of calreticulin might be involved in the low HLA-G surface expression on the 5-AC treated OCM-1A cells.
authors.affiliation: Laboratory Center, Wenzhou Medical College Affiliated Taizhou Hospital, Linhai 317000, China. yanwhcom@yahoo.com
authors.forename: Wei Hua
authors.initials: WH
authors.lastname: Yan
authors.forename: Ai Fen
authors.initials: AF
authors.lastname: Lin
authors.forename: Chien Chung
authors.initials: CC
authors.lastname: Chang
authors.forename: Soldano
authors.initials: S
authors.lastname: Ferrone
mesh_terms: D001374:Azacitidine / Q000031:analogs & derivatives* / Q000494:pharmacology; D037282:Calreticulin / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor; D002822:Choriocarcinoma / Q000378:metabolism*; D000077209:Decitabine; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006680:HLA Antigens / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008545:Melanoma / Q000378:metabolism*; D008745:Methylation; D011401:Promoter Regions, Genetic; D014158:Transcription, Genetic
chemical_list: D037282:Calreticulin; D006680:HLA Antigens; D059951:HLA-G Antigens; D015395:Histocompatibility Antigens Class I; D000077209:Decitabine; D001374:Azacitidine