医学分子生物学杂志 ›› 2022, Vol. 19 ›› Issue (1): 10-19.doi: 10.3870/j.issn.1672-8009.2022.01.002

• 论著 • 上一篇    下一篇

Hemin 诱导 K562 细胞分化前后 PU. 1 全基因组结合位点探测

  

  1. 上海海洋大学水产种质资源发掘与利用教育部重点实验室, 水产科学国家级实验教学示范中心 上海市, 201306
  • 出版日期:2022-01-31 发布日期:2022-02-25
  • 基金资助:
     国家自然科学基金面上项目 (No. 81770165) 

Genome-wide Identification of the Binding Sites of PU. 1 before and after Hemin-Induced Differentiation in K562 Cells

  1. Key Laboratory of Exploration and Utilization of Aquatic Germplasm Resources, Ministry of Education, Shanghai Ocean University; National Demonstration Center for Experimental Teaching of Aquatic Science, Shanghai Ocean University, Shanghai, 201306, China
  • Online:2022-01-31 Published:2022-02-25

摘要: 目的 探讨 K562 细胞分化前后全基因组上的 PU. 1 结合位点图谱及靶基因。 方法 用 hemin 对 K562 细胞诱导分化 72 h, 诱导分化前后的细胞利用 PU. 1 抗体进行染色质免疫共沉淀实验, 并进行高通量 测序, 利用生物信息学分析 K562 细胞分化前后 PU. 1 在基因组上的结合位点, 并进行基因注释、 基因功能 分析、 KEGG 通路分析及靶基因预测。 结果 发现 K562 细胞分化前 PU. 1 结合位点有 3 107 个, 分化后的 结合位点有 1 897 个, 共有的结合位点有 843 个。 KEGG 通路分析结果显示, 分化后 PU. 1 结合位点特有的 信号通路有 NF-κB 信号通路, 生长激素合成、 分泌和运输通路, MAPK 信号通路。 K562 细胞分化前后有 1 210个差异的 PU. 1 结合位点, 对应 179 个靶基因, 其中分化相关的 PU. 1 靶基因包括球蛋白基因、 自噬相 关基因、 microRNA、 非编码 RNA 以及转录因子。 Motif 分析结果显示, PU. 1 倾向于去结合 ACTTCC 特定序 列。 结论 PU. 1 的基因组结合位点在 K562 细胞分化前后发生了显著变化, 此研究结果为进一步揭示 PU. 1 在血细胞中的功能及作用机制提供了依据。 

关键词: K562 细胞, 分化, PU. 1, 染色质免疫共沉淀-高通量测序 

Abstract: Objective To examine the genome-wide identification of PU. 1 binding sites and target genes before and after hemin-induced differentiation in K562 cells. Methods K562 cells were treated with hemin for 72 h to induce cell differentiation. High-throughput sequencing was performed following chromatin immunoprecipitation with PU. 1 antibody in untreated and hemin-treated cell groups. The binding sites of PU. 1 in K562 cells before and after differentiation were analyzed by bioinformatics. Gene annotation, gene function analysis, KEGG pathway analysis and target genes prediction were performed. Results The results showed that there were 3 107 and 1 897 binding sites of PU. 1 before and after differentiation respectively. Among them, 843 binding sites were identified in both untreated and differentiated K562 cells. KEGG pathway analysis showed that the NF-κB signaling pathway, growth hormone synthesis, secretion and transport pathway, and MAPK signaling pathway were the unique signaling pathways related to PU. 1 binding after differentiation. Totally, 1 210 PU. 1 binding sites changed during differentiation, corresponding to 179 target genes. Moreover, the PU. 1 target genes related to differentiation included globin genes, autophagy-related genes, microRNA and non-coding RNA, and transcription factors. Motif analysis showed that PU. 1 tended to bind a specific ACTTCC sequence. Conclusion The genomic binding sites of PU. 1 changed significantly after hemin-induced differentiation in K562 cells. Our study provided a basis for further research on the function of PU. 1 in hematopoietic cell

Key words: K562 cell, differentiation, PU. 1, ChIP-seq

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