肿瘤干细胞中受H3K9me2调控的干性基因的筛选

Screening of Stem Genes Regulated by H3K9me2 in Tumor Stem Cells

为了分析比较甲基转移酶G9a和组蛋白H3K9me2修饰在胶质瘤干细胞与非干细胞中存在的差异,筛选出维持胶质瘤干细胞干性的相关基因。通过G9a抑制剂促进U87细胞成球和过表达G9a促进U87细胞分化的方法,培养了成球的胶质瘤干细胞和贴壁的非干细胞,这两种细胞的CD133表达差异明显。再利用H3K9me2抗体通过Ch IP-seq技术比较H3K9me2修饰在干细胞组与非干细胞组中的差异,在存在差异的基因中,对TSS±2 000 bp范围内的基因进行了GO分析,并随机选出10个转录因子进行QPCR验证,结果与Ch IP-seq实验基本一致。

In order to analyze and compare the modification differences of the methyltransferase G9 a and histone（H3K9me2）betweenin glioma stem cells and non-stem cells,the related genes maintaining the stem of glioma stem cells were screened. By using G9 a inhibitor tofacilitate U87 cells into spheres,and G9 a overexpression to promote the cell differentiation,the sphered glioma stem cells and monolayer nonstem cells were cultured,and there were significant differences in CD133 expression between the two cell models. Then we used H3K9me2 antibody to compare the differences of H3K9me2 modification between glioma stem cells and non-stem cells via Ch IP-seq assay. Genes in therange of TSS ± 2 000 bp among the genes with differences were selected for GO analysis. Moreover,10 transcription factors were randomlyselected to confirm the expression level by QPCR,and the result of which was almost consistent with that by Ch IP-seq assay.