DNA Methylation Profiles of Organic Anion Transporting Polypeptide 1B3 in Cancer Cell Lines
DNA Methylation Profiles of Organic Anion Transporting Polypeptide 1B3 in Cancer Cell Lines
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DOI：
10.1007/s11095-010-0064-3
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发表时间：
2010-03-01
影响因子：
3.7
通讯作者：
Sugiyama, Yuichi
中科院分区：
文献类型：
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作者：
Ichihara, Sayaka;Kikuchi, Ryota;Sugiyama, Yuichi
多特异性有机阴离子转运蛋白OATP 1B 3/SLCO 1B 3在多种癌细胞系和肿瘤组织中表达，其表达使细胞对某些抗癌药物敏感。本研究采用RT-PCR和亚硫酸氢盐测序法分别检测了OATP 1B 3基因转录起始位点（TSS）附近的DNA甲基化水平，并将其与肿瘤细胞和永生化细胞中OATP 1B 3基因mRNA的表达水平进行了相关性研究。和PK-45 P细胞，但在HepG 2、Caco-2和HEK 293细胞中低于检测限。亚硫酸氢盐测序表明，TSS周围的CpG二核苷酸在细胞系中甲基化程度不同，部分与OATP 1B 3的mRNA表达谱相关。此外，5-氮杂-2 '-脱氧胞苷（DNA甲基转移酶抑制剂）处理可使HepG 2和Caco-2细胞中OATP 1B 3的mRNA表达分别显著增加18倍和14倍，但在DLD-1和TFK-1细胞中则无此作用。DNA甲基化依赖性基因沉默至少部分参与了癌症/永生化细胞系中OATP 1B 3表达的调节。
Multispecific organic anion transporter, OATP1B3/SLCO1B3, is expressed in several cancer cell lines as well as tumor tissues, and its expression sensitizes the cells to some anti-cancer agents. The present study was aimed to characterize the DNA methylation profiles around the transcriptional start site (TSS) of OATP1B3 and correlate them with the mRNA expression in cancer and immortalized cell lines.The mRNA expression and DNA methylation profiles of OATP1B3 were determined by RT-PCR and bisulfite sequencing, respectively.The expression of OATP1B3 mRNA was detected in DLD-1, TFK-1, PK-8, and PK-45P cells, but below the limit of detection in HepG2, Caco-2, and HEK293 cells. Bisulfite sequencing demonstrated that CpG dinucleotides around the TSS are differentially methylated among cell lines and partly associated with the mRNA expression profile of OATP1B3. Furthermore, treatment with 5-aza-2'-deoxycytidine, an inhibitor of DNA methyltransferase, significantly increased the mRNA expression of OATP1B3 in HepG2 and Caco-2 cells by 18- and 14-fold, respectively, but not in DLD-1 and TFK-1 cells.DNA methylation-dependent gene silencing is at least partly involved in the regulation of OATP1B3 expression in cancer/immortalized cell lines.