DNA Methylation Suppresses Leptin Gene in 3T3-L1 Adipocytes.
DNA Methylation Suppresses Leptin Gene in 3T3-L1 Adipocytes.
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DOI：
10.1371/journal.pone.0160532
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Sakaue H
中科院分区：
文献类型：
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作者：
Kuroda M;Tominaga A;Nakagawa K;Nishiguchi M;Sebe M;Miyatake Y;Kitamura T;Tsutsumi R;Harada N;Nakaya Y;Sakaue H
瘦素是能量摄入和消耗的关键调节因子。这种肽激素在小鼠白色脂肪组织中表达，但在3 T3-L1脂肪细胞中几乎不表达。采用亚硫酸氢盐测序法，我们发现瘦素启动子中的CpG岛在3 T3-L1细胞中高度甲基化。5-氮杂胞苷，DNA甲基转移酶的抑制剂，显着增加瘦素的表达作为前脂肪细胞成熟为脂肪细胞。值得注意的是，瘦素的表达刺激胰岛素在脂肪细胞来源于前体细胞暴露于5-氮杂胞苷，但抑制噻唑烷二酮和地塞米松。相比之下，来自未经处理的前体细胞的脂肪细胞对5-氮杂胞苷和激素刺激均无反应，尽管在不存在去甲基化的情况下脂质积累足以促进瘦素表达。两者合计，结果表明，瘦素在3 T3-L1细胞中的表达需要脂肪形成前的DNA去甲基化，脂肪形成过程中的转录激活，以及脂肪形成后的脂质积累。
Leptin is a key regulator of energy intake and expenditure. This peptide hormone is expressed in mouse white adipose tissue, but hardly expressed in 3T3-L1 adipocytes. Using bisulfite sequencing, we found that CpG islands in the leptin promoter are highly methylated in 3T3-L1cells. 5-azacytidine, an inhibitor of DNA methyltransferase, markedly increased leptin expression as pre-adipocytes matured into adipocytes. Remarkably, leptin expression was stimulated by insulin in adipocytes derived from precursor cells exposed to 5-azacytidine, but suppressed by thiazolidinedione and dexamethasone. In contrast, adipocytes derived from untreated precursor cells were unresponsive to both 5-azacytidine and hormonal stimuli, although lipid accumulation was sufficient to boost leptin expression in the absence of demethylation. Taken together, the results suggest that leptin expression in 3T3-L1 cells requires DNA demethylation prior to adipogenesis, transcriptional activation during adipogenesis, and lipid accumulation after adipogenesis.