Kaiso 主要定位于体内的细胞核，与甲基化但非羟甲基化的 DNA 结合。
Kaiso mainly locates in the nucleus in vivo and binds to methylated, but not hydroxymethylated DNA.
机构信息
Beijing Key Laboratory of Carcinogenesis and Translational Research, Division of Cancer Etiology, Peking University Cancer Hospital & Institute, Beijing 100142, China.
出版信息
Chin J Cancer Res. 2015 Apr;27(2):148-55. doi: 10.3978/j.issn.1000-9604.2015.04.03.
OBJECTIVE
Kaiso is upregulated in many cancers and proposed to bind with both methylated- and unmethylated-DNA in the nucleus as a transcriptional repressor. The objective is to define its subcellular localization in vivo and exact binding DNA sequences in cells.
METHODS
Compartmentalization of exogenous Kaiso in cells was tracked with enhanced green fluorescence protein (EGFP) tag. The endogenous Kaiso expression in gastric carcinoma tissue was examined with immunohistochemical staining. Kaiso-DNA binding was tested using electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation assay (ChIP).
RESULTS
Kaiso mainly localized in the nucleus of cancer and stromal cells in vivo, but remained in the cytoplasm of cultured cells. Most importantly, nuclear Kaiso can bind with the methylated-CGCG-containing sequence in the CDKN2A promoter, but not with the hydroxymethylated-CGCG sequence in HCT116 cells.
CONCLUSIONS
Kaiso locates mainly in the nucleus in vivo where it binds with the methylated-CGCG sequences, but does not bind with the hydroxymethylated-CGCG sequences.
目的
Kaiso 在许多癌症中上调，并被提出作为转录抑制剂与核内甲基化和非甲基化的 DNA 结合。本研究旨在确定 Kaiso 在体内的亚细胞定位及其在细胞内的确切结合 DNA 序列。
方法
用增强型绿色荧光蛋白（EGFP）标记追踪外源性 Kaiso 的区室化。用免疫组织化学染色检测胃癌组织中内源性 Kaiso 的表达。用电泳迁移率变动分析（EMSA）和染色质免疫沉淀分析（ChIP）检测 Kaiso-DNA 结合。
结果
Kaiso 主要定位于体内癌细胞和基质细胞的核内，但在培养细胞的细胞质中仍存在。最重要的是，核 Kaiso 可以与 CDKN2A 启动子中含有甲基化-CGCG 的序列结合，但不能与 HCT116 细胞中羟甲基化-CGCG 序列结合。
结论
Kaiso 在体内主要定位于核内，与甲基化-CGCG 序列结合，但不与羟甲基化-CGCG 序列结合。
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