Analysis of Aberration for molecules of p53-depandent Apoptosis in hepatocellular carcinoma and Gastrointestinal Cancer

肝细胞癌和胃肠道癌中p53依赖性细胞凋亡分子的畸变分析

基本信息

批准号：
14570466
负责人：
NISHIDA Naoshi
金额：
$ 2.24万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14570466/
关键词：
Hepatocellular carcinoma P53 pathway p14 gene chromosomal instability deletion methylation carcinogenesis 突然変異発癌

项目摘要

We investigated mutation, deletion and aberrant methylation of the p14^<ARF> gene in 44 human hepatocellular carcinoma by using PCR-SSCP, direct sequencing, multiplex-PCR and methylation specific-PCR. In addition, we studied expression of p14^<ARF> with Real-Time PCR, expression of p53 with immunohistochemistry and examined chromosomalinstability by using semiquantitative microsatellite analysis. We found mutation of p14^<ARF> in two HCCs and deletion in one HCC, however aberrant promoter methylation was not detected in any cases.Examination of Real-Time PCR revealed that overexpression of p14^<ARF> was detected in all but one HCC with p14^<ARF> deletion, suggesting that aberration of transcriptional regulation was not take place during hepatocarcinogenesis. In addition, p14^<ARF> overexpression was associated with poorly differentiated phenotype. All three HCCs with p14^<ARF> alteration were well-differentiated. On the other hand, twelve HCCs showed aberration of p53 with immunohistochemistry and only one was well-differentiated HCC among them. These data indicate that p53 pathway play an important role in a subset of HCC formation.Chromosomal alteration was observed in all HCCs examined including well-differentiated HCC and p53 pathway aberration may accelerate chromosomal instability of HCC cell expanding altered chromosomal region and contribute to it's progression.

我们通过使用PCR-SSCP，直接测序，多重PCR和甲基化特异性PCR研究了44个人肝细胞癌中p14^<arf>基因的突变，缺失和异常甲基化。此外，我们研究了P14^<ARF>的表达，用实时PCR，具有免疫组织化学的p53表达，并通过使用半剂量的微卫星分析检查了染色体碱性。我们在两个HCC中发现了p14^<ARF>突变，并在一个HCC中缺失，但是在任何情况下均未检测到异常的启动子甲基化。实时PCR的检查表明，在所有情况下，p14^<arf> deletion的一个HCC中都检测到了p14^<arf>的过表达，该hcc在p14^<arf> deletion上进行了调查，该次数均在临床上进行了指示。另外，P14^<ARF>过表达与分化差的表型有关。所有三个具有P14^<ARF>改变的HCC均得到很好的差异。另一方面，十二个HCC表现出具有免疫组织化学的p53的像差，其中只有一个被很好地分化的HCC。这些数据表明，p53途径在HCC形成的子集中起着重要作用。在所检查的所有HCC中都观察到染色体改变，包括差异良好的HCC和p53途径的异常，可能会加速HCC细胞的染色体不稳定性，从而膨胀染色体区域，并促进染色体的进展。