In leukemia, knock-down of the death inducer-obliterator gene would inhibit the proliferation of endothelial cells by inhibiting the expression of CDK6 and CCND1

PeerJ. 2022 Feb 1:10:e12832. doi: 10.7717/peerj.12832. eCollection 2022.

Abstract

Background: 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.

Methods: 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.

Results: 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 death inducer-obliterator gene (DIDO) 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 DIDO showed a better overall survival (OS). The HUVEC cells were infected with shDIDO lentivirus, and reduced expression, inhibited proliferation, and increased apoptosis was observed in shDIDO cells. In addition, the expression of Cyclin-Dependent Kinase 6 (CDK6) and Cyclin D1 (CCND1) genes was inhibited in shDIDO cells. Finally, the public ChIP-seq data were used to analyze the regulators that bind with DIDO, and the H3K4me3 and PolII (RNA polymerase II) signals were found near the Exon1 and exon2 sites of DIDO.

Conclusion: The knock-down of DIDO will inhibit the proliferation of endothelial cells in the leukemia environment. The expression of DIDO may be regulated by H3K4me3 and the inhibition of DIDO may lead to the down-regulation of CDK6 and CCND1. However, how DIDO interacts with CDK6 and CCND1 requires further study.

Keywords: CCND1; CDK6; DIDO; Endothelial cells; Leukemia.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Proliferation / genetics
  • Cyclin D1* / genetics
  • Cyclin-Dependent Kinase 6 / genetics
  • Human Umbilical Vein Endothelial Cells / metabolism
  • Humans
  • Leukemia*

Substances

  • Cyclin D1
  • Cyclin-Dependent Kinase 6
  • CDK6 protein, human
  • CCND1 protein, human

Grants and funding

This study was supported by the National Natural Science Foundation of China (NO. 81360089), the Applied Basic Research in Yunnan Province of China (NO. 2019FE001-067, NO. 2017FE468-204), the Project of Educational Commission of Yunnan Province of China (NO. 2018JS229), the Applied Basic Research in Yunnan Province of China (NO. 202001AY070001-070), and the High-level Talents Training support program in Yunnan Province (YNWR-MY-2020-015). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.