Department of Hematology, The Third Affiliated Hospital of Kunming Medical University, Kunming, China.
Department of Orthopedics, Yan'an Hospital of Kunming City, The Affiliated Hospital of Kunming Medical University, Kunming, China.
PeerJ. 2022 Feb 1;10:e12832. doi: 10.7717/peerj.12832. eCollection 2022.
Endothelial cells (ECs) are a critical component of the hematopoietic niche, and the cross-talk between ECs and leukemia was reported recently. This study aimed to determine the genes involved in the proliferation inhibition of endothelial cells in leukemia.
Human umbilical vein endothelial cells (HUVEC) were cultured alone or co-cultured with K562 cell lines. GeneChip assays were performed to identify the differentially expressed genes. The Celigo, MTT assay, and flow cytometric analysis were used to determine the effect of RNAi DIDO on cell growth and apoptosis. The differently expressed genes were verified by qRT-PCR (quantitative real-time PCR) and western-blot.
In K562-HUVEC co-cultured cell lines, 323 down-regulated probes were identified and the extracellular signal-regulated kinase 5 (ERK5) signaling pathway was significantly inhibited. Among the down-regulated genes, the () is a part of the centrosome protein and may be involved in cell mitosis. As shown in the public data, leukemia patients with lower expression of showed a better overall survival (OS). The HUVEC cells were infected with lentivirus, and reduced expression, inhibited proliferation, and increased apoptosis was observed in cells. In addition, the expression of () and () genes was inhibited in cells. Finally, the public ChIP-seq data were used to analyze the regulators that bind with , and the H3K4me3 and PolII (RNA polymerase II) signals were found near the Exon1 and exon2 sites of .
The knock-down of will inhibit the proliferation of endothelial cells in the leukemia environment. The expression of may be regulated by H3K4me3 and the inhibition of may lead to the down-regulation of and . However, how interacts with and requires further study.
内皮细胞（ECs）是造血龛的关键组成部分，最近有报道称 ECs 与白血病之间存在相互作用。本研究旨在确定参与白血病中内皮细胞增殖抑制的相关基因。
单独培养或与 K562 细胞系共培养人脐静脉内皮细胞（HUVEC）。采用基因芯片技术鉴定差异表达基因。Celigo 细胞计数仪、MTT 检测法和流式细胞术分析 RNAi DIDO 对细胞生长和凋亡的影响。采用 qRT-PCR（实时定量 PCR）和 Western blot 验证差异表达基因。
在 K562-HUVEC 共培养细胞系中，鉴定出 323 个下调探针，其中细胞外信号调节激酶 5（ERK5）信号通路受到显著抑制。在下调基因中，（）是中心体蛋白的一部分，可能参与细胞有丝分裂。公共数据显示，表达水平较低的白血病患者总体生存率（OS）更好。用慢病毒感染 HUVEC 细胞，观察到下调 基因后细胞增殖受到抑制，凋亡增加。此外，下调 基因会抑制（）和（）基因的表达。最后，利用公共 ChIP-seq 数据分析与 结合的调控因子，发现 H3K4me3 和 PolII（RNA 聚合酶 II）信号位于 外显子 1 和外显子 2 附近。
敲低 基因会抑制白血病环境中内皮细胞的增殖。 基因的表达可能受 H3K4me3 调控，而 基因的抑制可能导致 和 基因下调。然而，与 基因相互作用的确切机制还需要进一步研究。
