Discovery of epigenetically masked tumor suppressor genes in gyneoobgical cancer

发现妇科癌症中表观遗传屏蔽的抑癌基因

• 批准号: 18591840
• 负责人: TAKAI Noriyuki
• 金额: $ 2.44万
• 依托单位: Oita University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2006
• 资助国家: 日本
• 起止时间: 2006 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18591840/
• 关键词: Epigenetic; Histone; Microarray; Methvlation; Tumor suppressor gene; Gynecologic cancer; ヒストン修飾

Realization that many tumor suppressor genes are silenced by epigenetic mechanisms has stimulated discovery of novel tumor suppressor genes. We used a variety of research tools to search for genes that are epigenetically silenced in human endometrial cancers. Changes in global gene expression of the endometrial cancer cell line Ishikawa was analyzed after treatment with the demethylating agent 5-aza-2'-deoxycytidine (5-Aza-CdR) combined with the histone deacetylase inhibitor, suberoyl anilide bishydroxamide (SAHA). By screening over 22,000 genes, candidate tumor suppressor genes were identified. Additional microarray analysis and real-time RTPCR of normal and cancerous endometrial samples, and search for CpG islands further refined the list. Tigl and C/ebpa were chosen for further study. Expression of both genes was low in endometrial cancer cell lines and clinical samples, but high in normal endometrial tissues. Bisulfite sequencing and restriction analysis revealed aberrant methylation of the CpG island in the Tigl gene of all 6 endometrial cancer cell lines examined, whereas the C/ebpa promoter remained unmethylated in endometrial cancers Chromatin immunoprecipitation showed increased acetylated histones H3 bound to both Tigl and C/ebpa genes after treatment with 5-Aza-CdR and/or SAHA. Forced expression of either TIG1 or C/EBPa led to significant growth reduction of Ishikawa cells. Immunoprecipitation and reporter gene assays demonstrated that C/EBPa bound to and suppressed transcriptional activity of E2F1, a key cell cycle regulator. Our data suggest that C/ebpa and Tigl function as tumor suppressor proteins in endometrial cancers and that their reexpression may be a therapeutic target.

许多肿瘤抑制基因被表观遗传机制沉默的认识刺激了新型肿瘤抑制基因的发现。我们使用了各种研究工具来寻找在人类子宫内膜癌中表观遗传沉默的基因。用去甲基化剂5-氮杂-2 '-脱氧胞苷（5-Aza-CdR）联合组蛋白去乙酰化酶抑制剂辛二酰苯胺双羟酰胺（SAHA）处理子宫内膜癌细胞系石川后，分析其整体基因表达的变化。通过筛选超过22，000个基因，确定了候选肿瘤抑制基因。对正常和癌性子宫内膜样本的额外微阵列分析和实时RTPCR，以及对CpG岛的搜索进一步完善了该列表。选择Tigl和C/ebpa进行进一步研究。两种基因在子宫内膜癌细胞系和临床标本中均呈低表达，而在正常子宫内膜组织中呈高表达。亚硫酸氢盐测序和限制性酶切分析显示，所有6个子宫内膜癌细胞系的Tigl基因中的CpG岛异常甲基化，而C/ebpa启动子在子宫内膜癌中保持未甲基化。染色质免疫沉淀显示，用5-Aza-CdR和/或SAHA处理后，与Tigl和C/ebpa基因结合的乙酰化组蛋白H3增加。TIG 1或C/EBPa的强制表达导致石川细胞的显著生长减少。免疫沉淀和报告基因分析表明，C/EBPa结合并抑制转录活性的E2 F1，一个关键的细胞周期调节因子。我们的数据表明，C/ebpa和Tigl作为子宫内膜癌的肿瘤抑制蛋白的功能，它们的重新表达可能是一个治疗靶点。

项目成果

M344 is a novel synthesized histone deacetylase inhibitor that induces growth inhibition, cell cycle arrest, and apoptosis in human endometrial cancer and ovarian cancer cells.

• 发表时间: 2006
• 期刊: Gynecologic oncology
• 影响因子: 4.7
• 作者: N. Takai;T. Ueda;M. Nishida;K. Nasu;H. Narahara

K252a is highly effective in suppressing the growth of human endometrial cancer cells,but has little effect on normal human endometrial epithelial cells

K252a对抑制人子宫内膜癌细胞的生长非常有效，但对正常人子宫内膜上皮细胞影响不大

• 发表时间: 2008
• 期刊: Oncol Rep 19
• 作者: Kiyotake;Ichizuka;Noriyuki Takai;Noriyuki Takai;Noriyuki Takai
• 通讯作者: Noriyuki Takai

エピジェネティクスによる婦人科腫瘍への治療と新しい癌抑制遺伝子

利用表观遗传学和新抑癌基因治疗妇科肿瘤

• 发表时间: 2008
• 作者: Noriyuki;Takai;Noriyuki Takai;Noriyuki Takai;高井 教行
• 通讯作者: 高井 教行

A case pericarditis carcinomatosa arising torn cervical cancer

宫颈癌撕裂所致癌性心包炎一例

• 发表时间: 2007
• 作者: Naoko;Kira
• 通讯作者: Kira

過多月経・月経困難症に対するレボノルゲストレル徐放性IUSの有効性

左炔诺孕酮缓释 IUS 治疗月经过多/痛经的疗效

• 发表时间: 2008
• 期刊: 産科と婦人科 75
• 作者: Kiyotake;Ichizuka;Noriyuki Takai;Noriyuki Takai;Noriyuki Takai;Noriyuki Takai;Kaei Nasu;Kaei Nasu;高井 教行
• 通讯作者: 高井 教行