前期工作发现OSKM介导C3A人肝癌细胞重编程,获诱导多潜能肿瘤干细胞C3A-iPCs,CD44表达并核转位,内源性重编程基因表达,保持肿瘤干细胞Primed状态。Crispr方法敲除CD44基因,获得CD44KO-C3A-iPCs细胞重编程基因的表达大幅度提高,提示保持肿瘤干细胞Naive-like重编程状态。本项目将研究两种不同状态的细胞与转录调节和表观遗传记忆机制。进一步定向肝诱导分化,拟研究Naive-like重编程状态的诱导肿瘤干细胞的转归。证明CD44基因携带表观遗传记忆,Primed状态的诱导多潜能肿瘤干细胞没有擦除肿瘤“自已”的表观标志,而Naive-like重编程状态CD44的表观遗传标志被擦除。本研究将揭示诱导多潜能肿瘤干细胞的Primed和Naive表观遗传状态可在特定的条件下互相转换,与DNA甲基化和组蛋白修饰机制相关。研究将为临床肿瘤治疗提出新的研究思路和前景。

Based on previous work found OSKM-mediated reprogramming of C3A human hepatoma cells, induced pluripotent cancer stem-like cells (C3A-iPCs) expressed endogenous reprogramming genes, expressed CD44 and nuclear translocation, maintained “primed” state of iPCs. CD44KO-C3A-iPCs were obtained with Crispr-cas9-mediated CD44 gene knockout in C3A-iPCs, expressed endogenous reprogramming genes in a substantial increase and maintained "naive" –like state of iPCs. Both primed and naive-like states of iPCs we studies its transcriptional regulation and epigenetic memory mechanism. Further, directed hepatic differentiation we will research these cells how to be turned back in naive-like reprogramming state of iPCs. Herein we will study CD44 may be carried the epigenetic memory and does not erase the tumor"self" epigenetic markings in primed state, but in a naive-like reprogramming state CD44 epigenetic information is erased. Expected results both primed and naive states of iPCs may be converted to each other under certain conditions, associated with DNA methylation and histone modification mechanism. The study will propose new ideas and perspectives in clinical tumor therapy.