Epigenetic up-regulation of c-c chemokine receptor 7 and C-X-C chemokine receptor 4 expression in melanoma cells
Epigenetic up-regulation of c-c chemokine receptor 7 and C-X-C chemokine receptor 4 expression in melanoma cells
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DOI：
10.1158/0008-5472.can-04-3531
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发表时间：
2005-03-01
期刊：
影响因子：
11.2
通讯作者：
Hoon, DSB
中科院分区：
文献类型：
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作者：
Mori, T;Kim, J;Hoon, DSB
组蛋白去乙酰化和DNA甲基化建立了表观遗传修饰，通过染色质重塑可能导致基因沉默。我们假设黑色素瘤细胞表面的趋化因子受体C-C趋化因子受体7(CCR7)和C-X-C趋化因子受体4(CXCR4)经历了表观遗传调控。我们研究了组蛋白脱乙酰酶抑制剂和去甲基剂是否影响黑色素瘤细胞CCR7和CXCR4的表达。首先，进行基因芯片分析以筛选曲古菌素A(TSA)和5-氮杂-2-脱氧胞苷(5-Aza)处理黑色素瘤细胞后趋化因子受体表达的变化。CCR7和CXCR4的mRNA表达均发生改变，并选择作进一步研究。采用实时定量逆转录-聚合酶链式反应、免疫组织化学和Western印迹方法检测TSA和5-aza诱导的黑色素瘤细胞的mRNA和蛋白表达的变化。细胞迁移实验评估CCR7和CXCR4表达改变对细胞功能的影响。经TSA或5-aza处理后，黑色素瘤细胞CCR7和CXCR4的基因表达均增加。TSA是最强的促进剂。联合用药后，CCR7和CXCR4的mRNA表达也上调。联合处理后的免疫组织化学显示CCR7和CXCR4的染色均较对照细胞增强。在TSA和5-aza处理细胞中，CCR7和CXCR4的黑色素瘤细胞迁移率分别是对照细胞的2倍。综上所述，组蛋白脱乙酰酶抑制剂和去甲基剂上调了黑色素瘤细胞上CCR7和CXCR4的表达。趋化因子受体表达的增加与功能活动相关。最重要的是，我们已经确定了一种表观遗传学机制，它可能内源性地调节黑色素瘤细胞上趋化因子受体的表达。
Histone deacetylation and DNA methylation establish epigenetic modifications, which through chromatin remodeling may result in gene silencing. We hypothesized that chemokine receptors C-C chemokine receptor 7 (CCR7) and C-X-C chemokine receptor 4 (CXCR4) on melanoma cells undergo epigenetic regulation. We investigated whether a histone deacetylase inhibitor and a demethylating agent influence CCR7 and CXCR4 expression on melanoma cells. Initially, microarray analysis was done to screen changes in chemokine receptor expression on melanoma cells after treatment with trichostatin A (TSA) and 5-Aza-2-deoxycytidine (5-Aza). CCR7 and CXCR4 mRNA expression were uniformly altered and selected for further investigation. Quantitative real-time reverse transcription-PCR assay, immunohistochemistry, and Western blot analysis were used to assess changes in mRNA and protein expression induced by TSA and 5-Aza in melanoma lines. Cell migration assays were conducted to assess the effects of altered CCR7 and CXCR4 expression on cell function. Treatment with TSA or 5-Aza increased gene expression of both CCR7 and CXCR4 in melanoma lines. TSA was the strongest enhancer. With combined treatment, CCR7 and CXCR4 mRNA expression was also up-regulated. Immunohistochemistry after combined treatment showed enhanced staining of both CCR7 and CXCR4 compared with control cells. Melanoma cell migration in TSA and 5-Aza-treated cells was land 2-fold higher than control cells for CCR7 and CXCR4, respectively. In summary, a histone deacetylase inhibitor and a demethylating agent up-regulated CCR7 and CXCR4 expression on melanoma cells. This increase in chemokine receptor expression correlated with functional activity. Most importantly, we have identified an epigenetic mechanism that may endogenously regulate chemokine receptor expression on melanoma cells.