Modulation by decitabine of gene expression and growth of osteosarcoma U2OS cells in vitro and in xenografts: identification of apoptotic genes as targets for demethylation.
Modulation by decitabine of gene expression and growth of osteosarcoma U2OS cells in vitro and in xenografts: identification of apoptotic genes as targets for demethylation.
复制标题
根据基因表达和骨肉瘤U2OS细胞在体外和异种移植中的生长的调节：凋亡基因作为脱甲基化的靶标。
DOI：
10.1186/1475-2867-7-14
复制
发表时间：
2007-09-10
影响因子：
5.8
通讯作者：
Squire, Jeremy A.
中科院分区：
文献类型：
--
作者：
Al-Romaih, Khaldoun;Somers, Gino R.;Bayani, Jane;Hughes, Simon;Prasad, Mona;Cutz, Jean-Claude;Xue, Hui;Zielenska, Maria;Wang, Yuzhuo;Squire, Jeremy A.
甲基化介导的基因沉默是与癌症有关的一种表观遗传机制。关于利用 DNA 甲基化抑制剂（例如地西他滨）在骨肉瘤 (OS) 中调节基因表达的作用的研究还很有限。从生物学角度理解地西他滨对 OS 的总体影响非常重要，因为这种特殊药物目前正在进行临床试验。本研究的目的是测量 OS 细胞系 U2OS 对体外和体内地西他滨治疗的反应。微阵列表达谱用于区分 U2OS 细胞中地西他滨依赖性基因表达变化，并鉴定具有去甲基化 CpG 启动子区域的响应位点。在免疫缺陷小鼠的肾被膜下建立U2OS异种移植物，以研究地西他滨在体内对肿瘤生长和分化的影响。通过使用 5-甲基胞苷抗体的免疫组织化学，可以在来自治疗小鼠的异种移植物中检测到核甲基化水平降低。地西他滨治疗显着减小了肿瘤异种移植物的大小（p < 0.05）。处理后的 U2OS 异种移植物切片的组织学分析显示，有丝分裂活性较低（p < 0.0001），骨基质生成增加（p < 0.0001），并且凋亡细胞数量较多（p = 0.0329）。 U2OS 培养细胞的微阵列表达谱显示，地西他滨治疗引起 88 个基因表达的显着诱导（p < 0.0025）。 13 个有 ≥2 倍的变化，其中 11 个具有 CpG 岛相关启动子。有趣的是，这 11 个基因中的 6 个是促凋亡基因，地西他滨在体外显着诱导 U2OS 细胞死亡（p < 0.05）。 6 个促凋亡基因（GADD45A、HSPA9B、PAWR、PDCD5、NFKBIA 和 TNFAIP3）在体内也被诱导至 ≥2 倍。定量甲基化焦磷酸测序证实，由于 U2OS 地西他滨在体外和异种移植物中的治疗，测试的促凋亡基因具有 CpG 岛 DNA 去甲基化。这些数据提供了关于在 OS 中使用表观遗传修饰剂的新见解，并对涉及去甲基化药物的治疗试验具有重要意义。总的来说，这些数据提供了生物学证据，表明地西他滨的一种作用模式可能是利用 OS 细胞死亡途径中特定效应子的启动子 CpG 去甲基化诱导细胞凋亡。
Methylation-mediated silencing of genes is one epigenetic mechanism implicated in cancer. Studies regarding the role of modulation of gene expression utilizing inhibitors of DNA methylation, such as decitabine, in osteosarcoma (OS) have been limited. A biological understanding of the overall effects of decitabine in OS is important because this particular agent is currently undergoing clinical trials. The objective of this study was to measure the response of the OS cell line, U2OS, to decitabine treatment both in vitro and in vivo. Microarray expression profiling was used to distinguish decitabine-dependent changes in gene expression in U2OS cells, and to identify responsive loci with demethylated CpG promoter regions. U2OS xenografts were established under the sub-renal capsule of immune-deficient mice to study the effect of decitabine in vivo on tumor growth and differentiation. Reduced nuclear methylation levels could be detected in xenografts derived from treated mice by immunohistochemistry utilizing a 5-methylcytidine antibody. Decitabine treatment reduced tumor xenograft size significantly (p < 0.05). Histological analysis of treated U2OS xenograft sections revealed a lower mitotic activity (p < 0.0001), increased bone matrix production (p < 0.0001), and a higher number of apoptotic cells (p = 0.0329). Microarray expression profiling of U2OS cultured cells showed that decitabine treatment caused a significant induction (p < 0.0025) in the expression of 88 genes. Thirteen had a ≥2-fold change, 11 of which had CpG-island-associated promoters. Interestingly, 6 of these 11 were pro-apoptotic genes and decitabine resulted in a significant induction of cell death in U2OS cells in vitro (p < 0.05). The 6 pro-apoptotic genes (GADD45A, HSPA9B, PAWR, PDCD5, NFKBIA, and TNFAIP3) were also induced to ≥2-fold in vivo. Quantitative methylation pyrosequencing confirmed that the tested pro-apoptotic genes had CpG-island DNA demethylationas a result of U2OS decitabine treatment both in vitro and in xenografts These data provide new insights regarding the use of epigenetic modifiers in OS, and have important implications for therapeutic trials involving demethylation drugs. Collectively, these data have provided biological evidence that one mode of action of decitabine may be the induction of apoptosis utilizing promoter-CpG demethylation of specific effectors in cell death pathways in OS.
登录
查看更多内容
影响因子：
11.2
作者：
Eramo, A;Pallini, R;De Maria, R
通讯作者：
De Maria, R
影响因子：
2.8
作者：
Doxsee, Daniel W.;Gout, Peter W.;Wang, Yu-Zhuo
通讯作者：
Wang, Yu-Zhuo
影响因子：
64.8
作者：
Barreto, Guillermo;Schaefer, Andrea;Niehrs, Christof
通讯作者：
Niehrs, Christof
影响因子：
11.5
作者：
Cutz, Jean-Claude;Guan, Jun;Wang, Yu-Zhuo
通讯作者：
Wang, Yu-Zhuo
影响因子：
8
作者：
Karpf, AR;Jones, DA
通讯作者：
Jones, DA