THE DEVELOPMENT OF A GENE-BASED DIAGNOSTIC METHOD TO EVALUATE THE EFFICACY OF PRE-CHEMOTHERAPUTIC AGENTS IN ORAL CANCER THAT ALTER GENE EXPRESSION BY AFFECTING DNA METHYLATION

开发基于基因的诊断方法来评估通过影响 DNA 甲基化来改变基因表达的口腔癌化疗前药物的疗效

• 批准号: 15592121
• 负责人: NOGUCHI Makoto
• 金额: $ 2.24万
• 依托单位: SAPPORO MEDICAL UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15592121/
• 关键词: oral cancer; pre-chemotheraputic agents; sensitivity of chemotherapy; DNA methylation; docetaxel; RNAi

(1)In oral cancer cell (OCC) lines, DNA methylation of the CHFR gene was examined as a target to evaluate the sensitivity of pre-chemotherapeutic agents.(2)To examine the role of CHFR in the prophase mitotic checkpoint in more detail, we used cells which do not express CHFR to evaluate the mitotic index after treatment of microtubule inhibitors such as docetaxel. There is a significant correlation between the absence of CHFR expression and a high mitotic index in OCC lines. Re-expression of CHFR, by blocking promoter methylation through drug treatment (i.e. 5-aza-dC) reduced the mitotic index in cells. This indicates that CHFR regulates a checkpoint importance in controlling entry into mitosis. Thus, cells lacking CHFR apparently do not stop at prophase and enter mitosis when treated with microtubule inhibitors.(3)In addition to oral cancer, abnormality of CHFR was observed in colon cancer, stomach cancer, and leukemia. To verify that the sensitivity of CHFR-deficient OCC to docetaxel is specific to microtubule inhibitors, cells were treated will another microtubule inhibitor (paclitaxel), a topoisomerase inhibitor (VP16), or an alkylating agent (CDDP). Some cells were sensitive to paclitaxel but not to VP16 or CDDP, indicating that the CHFR checkpoint is involved in the response to mitotic stress but not in the response to topoisomerase inhibition or double strand breaks.(4)Disruption of CHFR by short hairpin RNA reduced the ability of cells to regulate G2/M phase transition due to impaired mitotic checkpoint. In addition, CHFR knockdown in OCC increases sensitivity of the cells to microtubule inhibitors. Our results indicate that CHFR can be a molecular target for chemotherapy.

(1)在口腔癌细胞（OCC）系中，检测CHFR基因的DNA甲基化作为评价化疗前药物敏感性的靶点。(2)为了更详细地研究CHFR在前期有丝分裂检查点中的作用，我们使用不表达CHFR的细胞来评估微管抑制剂（如多西他赛）治疗后的有丝分裂指数。在OCC细胞系中，CHFR表达缺失与高有丝分裂指数之间存在显著相关性。通过药物治疗（即5-aza-dC）阻断启动子甲基化，CHFR的重新表达降低了细胞中的有丝分裂指数。这表明CHFR调节检查点在控制有丝分裂进入中的重要性。因此，缺乏CHFR的细胞在接受微管抑制剂治疗时，显然不会在前期停止并进入有丝分裂。(3)除口腔癌外，CHFR在结肠癌、胃癌、白血病中均有异常。为了验证chfr缺陷OCC对多西紫杉醇的敏感性是微管抑制剂特异性的，我们将细胞用另一种微管抑制剂（紫杉醇）、拓扑异构酶抑制剂（VP16）或烷基化剂（CDDP）处理。一些细胞对紫杉醇敏感，但对VP16或CDDP不敏感，这表明CHFR检查点参与了对有丝分裂应激的反应，而不是对拓扑异构酶抑制或双链断裂的反应。(4)由于有丝分裂检查点受损，短发夹RNA破坏CHFR降低了细胞调节G2/M期转变的能力。此外，OCC中CHFR的下调增加了细胞对微管抑制剂的敏感性。我们的研究结果表明CHFR可以成为化疗的分子靶点。

项目成果

Small interfering RNA-Induced CHFR silencing sensitizes oral squamous cell cancer cells to microtubule inhibitors

• DOI: 10.4161/cbt.4.7.1896
• 发表时间: 2005-07-01
• 期刊: CANCER BIOLOGY & THERAPY
• 影响因子: 3.6
• 作者: Ogi, K;Toyota, M;Tokino, T
• 通讯作者: Tokino, T

Small interfering RNA-induced CHFR silencing sensitizes oral squamous cell cancer cells to microtubule inhibitots.

小干扰 RNA 诱导的 CHFR 沉默使口腔鳞状细胞癌细胞对微管抑制剂敏感。

• 发表时间: 2005
• 期刊: Cancer Biol Ther. 4(7)
• 作者: Ogi K;Toyota M;Mita H;Satoh A;Kashima L;Sasaki Y;Suzuki H;Nishikawa N;Noguchi M;Shinomura Y;Hiratsuka H;Tokino T
• 通讯作者: Tokino T