SPAN-Xb expression in myeloma cells is dependent on promoter hypomethylation and can be upregulated pharmacologically
SPAN-Xb expression in myeloma cells is dependent on promoter hypomethylation and can be upregulated pharmacologically
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DOI：
10.1002/ijc.21499
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发表时间：
2006-03-15
影响因子：
6.4
通讯作者：
Lim, SH
中科院分区：
文献类型：
--
作者：
Wang, ZQ;Zhang, J;Lim, SH
SPAN-Xb是多发性骨髓瘤（MM）中的一种新的癌-睾丸抗原。在这项研究中，我们确定的机制，调节斯潘-Xb在MM中的表达。斯潘-Xb启动子序列克隆到CAT报告载体，以确定启动子甲基化在基因表达调控中的作用。用5-氮杂胞苷处理肿瘤细胞，并使用一组细胞因子来确定它们诱导Span-Xb表达的能力。将亚硫酸氢盐转化与序列分析应用于一组肿瘤细胞和正常组织，以关联CpG二核苷酸低甲基化和SPAN-Xb表达。我们发现，甲基化可以沉默SPAN-Xb启动子功能。5-氮杂胞苷诱导启动子低甲基化，并导致在转录本和蛋白质水平的SPAN-Xb的表达。在SPAN-Xb启动子内的位置-310、-307、-299和-221处的CpG二核苷酸的低甲基化强烈预测SPAN-Xb表达。IL-7和GM-CSF也能够上调骨髓瘤细胞中的SPAN-Xb的表达，但仅在启动子序列被低甲基化之后。我们的结果提供了第一个证据，表明启动子甲基化在斯潘-Xb的初级调节中的作用以及IL-7和GM-CSF进一步增强骨髓瘤细胞中斯潘-Xb基因和蛋白表达的能力。(c)2005 Wiley-Liss，Inc.
SPAN-Xb is a novel cancer-testis antigen in multiple myeloma (MM). In this study, we determined the mechanisms regulating SPAN-Xb expression in MM. SPAN-Xb promoter sequence was first cloned into the CAT-reporter vector to determine the role of promoter methylation in the regulation of gene expression. Tumor cells were treated with 5-azacytidine and a panel of cytokines were used to determine their ability to induce SPAN-Xb expression. Bisultite conversion with sequence analysis was applied to a panel of tumor cells and normal tissues to correlate the CpG dinucleotide hypomethylation and SPAN-Xb expression. We found that SPAN-Xb promoter function could be silenced by methylation. 5-Azacytidine induced promoter hypomethylation and resulted in SPAN-Xb expression, at both the transcript and protein levels. Hypomethylation of the CpG dinucleotides at positions -310, -307, -299 and -221 within the SPAN-Xb promoter strongly predict for SPAN-Xb expression. Both IL-7 and GM-CSF were also able to upregulate the expression of SPAN-Xb in myeloma cells, but only after the promoter sequence has been hypomethylated. Our results provide the first evidence showing the role of promoter methylation in the primary regulation of SPAN-Xb and the ability of IL-7 and GM-CSF to further enhance SPAN-Xb gene and protein expression in myeloma cells. (c) 2005 Wiley-Liss, Inc.