The allelic expressiondifference of humanMDM2gene in endometrial cancer

人类MDM2基因在子宫内膜癌中的等位基因表达差异

• 批准号: 23791847
• 负责人: TSUNEMATSU Ryosuke
• 金额: $ 2.66万
• 依托单位: Kyushu University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23791847/
• 关键词: AED; 子宮体癌; MDM2; PCR-SSCP

#1. To search the AEDs of human MDM2gene, we analyzed 45 samples from the endometrial cancer patients and the healthy controls by PCR-SSCP-based method. Out of 20 endometrial cancer patients, we identified two cases presenting AED. (fig.1) #2. We confirmed that these AED derived from P2 promotor of MDM2gene. And we found new SNP (T/C) on the 5’-upstream of the SNP309 in the MDM2P2 promotor. We found that this SNP created an NF-・B consensus sequence in the Sp1 binding sequence. #3. Using the Chromatin Immunoprecipitation (ChIP) and the luciferase reported assays, we confirmed that the NF-・B consensus sequence formedby the newly-discovered SNP(C-allele) bound to NF-・B, but not T-allele, and that NF-・B can suppress the transcription of MDM2gene. #4. Immnohistochemistry analysis showed that NF-・??p50 protein accumulated in the nuclei of the AED-positive endometrial cancer tissue. #5. The Biacore analysis revealed that p50 preferred C-allele to T-allele, conversely, Sp1 T-allele to C-allele. We also found that they bound to the same DNA region in a competitive manner.

#1.为寻找人类MDM 2基因的抗癫痫药物，采用PCR-SSCP方法对45例子宫内膜癌患者和45例健康对照进行了分析。在20例子宫内膜癌患者中，我们确定了两例AED。（图1）#2.我们证实这些AED均来源于MDM 2基因的P2启动子。在MDM 2 P2启动子SNP 309的5 '上游发现了新的SNP（T/C）。我们发现该SNP在Sp1结合序列中产生NF-·B共有序列。#3.通过染色质免疫沉淀（ChIP）和荧光素酶报告试验，证实了新发现的SNP（C等位基因）形成的NF-·B共有序列与NF-·B结合，而与T等位基因不结合，NF-·B对MDM 2基因的转录有抑制作用。#4.免疫组化结果显示NF-？p50蛋白在AED阳性的子宫内膜癌组织细胞核内聚集。#5. Biacore分析显示，p50基因对C等位基因的偏好大于T等位基因，而Sp1基因对T等位基因的偏好大于C等位基因。我们还发现它们以竞争的方式结合到同一DNA区域。