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PCR-SSCP、直接测序、多重PCR和甲基化特异性PCR研究了44例人肝细胞癌中p14^<ARF>基因的突变、缺失和异常甲基化。此外，我们用实时PCR研究了p14^<ARF>的表达，用免疫组织化学研究了p53的表达，并通过半定量微卫星分析检查了染色体稳定性。我们在两个 HCC 中发现 p14^<ARF> 突变，在一个 HCC 中发现 p14^<ARF> 缺失，但在任何情况下均未检测到异常启动子甲基化。实时 PCR 检查显示，除 1 个 HCC 之外，在所有 HCC 中均检测到 p14^<ARF> 过表达具有 p14^<ARF> 缺失的 HCC，表明在肝癌发生过程中并未发生转录调控的异常。此外，p14^<ARF>过表达与低分化表型相关。所有三种具有 p14^<ARF> 改变的 HCC 均分化良好。另一方面，免疫组化显示 12 例 HCC 存在 p53 异常，其中只有 1 例是分化良好的 HCC。这些数据表明，p53 通路在 HCC 形成的一个子集中发挥着重要作用。在所有检查的 HCC（包括分化良好的 HCC）中均观察到染色体改变，p53 通路畸变可能会加速 HCC 细胞的染色体不稳定，扩大改变的染色体区域并有助于其进展。