Expression of MART-1 in human uveal melanoma cell lines / 中华眼底病杂志

ABSTRACT

Objective To observe the expression and transcription of MART-1 in human uveal melanoma cell lines 92-1,92-2,Ocm3,Me1285,as well as the possible effect ofmethylation on its expression.Methods The cell lines 92-1,92-2,Ocm3 and Me1285 were cultured routinely and tested for MART-1 expression at protein and mRNA level by FACS analysis,Western blot and RT-PCR respectively.Methylation status of the MART-1 promoter region in all the cell lines were checked by Southern blots of DNA digested with methylation sensitive restriction enzymes.Results As observed in FACS analysis and Western blot,92-1,92-2 and Ocm3 were MART-1 positive cell lines while Me1285 was negative cell line.Consistent with protein analysis,92-1 and Ocm3 cell lines showed MART-1 specific PCR products and there was no product in Me1285 cell line in RT-PCR.The MART-1 positive cell lines,92-1,92-2,and Ocm3 show methylation at the MspⅠ/Hpa Ⅱ site,and the Nru Ⅰ sites of all positive cell lines are not methylated.The MART-1 negative cell line Me1285 shows hyperrnethylation at the Nru Ⅰ site and the Msp Ⅰ/Hpa Ⅱ site is not methylated.Conclusions MART-1 could be expressed in human uveal melanoma cell lines 92-1,92-2 and Ocm3.The change of methylation status of MART-1 promoter may correlate with the transcription of MART-1.