Delineation of EZH2 oncogenic functions in hepatocellular carcinoma. Homo sapiens

The goal of this study was to delineate the important EZH2 direct target genes that mediate the oncogenic properties of EZH2 in HCC. The EZH2 direct target genes in two HCC cell lines were identified by chromatin immunoprecipitation microarray (ChIP-chip) analysis and later confirmed by independent ChIP-PCR. The functions of the target genes were further examined. Overall design: Two human HCC cell lines, Huh7 and PLC/PRF/5 (PLC5), were grown in DMEM supplemented with 10% fetal bovine serum. ChIP assays were performed using anti-EZH2 antibody. The immunoprecipitated and input DNA were used to probe the Agilent human ChIP-chip arrays.

本研究旨在阐明介导肝细胞癌（Hepatocellular Carcinoma, HCC）中EZH2致癌特性的关键直接靶基因。通过染色质免疫沉淀微阵列（ChIP-chip）分析，鉴定了两株人肝细胞癌（HCC）细胞系中的EZH2直接靶基因，并通过独立的染色质免疫沉淀PCR（ChIP-PCR）实验完成验证。随后对这些靶基因的功能开展了进一步研究。
整体实验设计：采用添加10%胎牛血清的达尔伯克改良伊格尔培养基（DMEM）培养两株人源肝细胞癌（HCC）细胞系Huh7与PLC/PRF/5（简称PLC5）。使用抗EZH2抗体进行染色质免疫沉淀实验，将免疫沉淀所得DNA与输入对照DNA用于安捷伦人源ChIP-chip芯片的杂交检测。