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
Honghua Cao,
Lilan Wang,
Chengkui Geng,
Man Yang,
Wenwen Mao,
Linlin Yang,
Yin Ma,
Ming He,
Yeying Zhou,
Lianqing Liu,
Xuejiao Hu,
Jingxing Yu,
Xiufen Shen,
Xuezhong Gu,
Liefen Yin,
Zhenglei Shen
Affiliations
Honghua Cao
Department of Hematology, The Third Affiliated Hospital of Kunming Medical University, Kunming, China
Lilan Wang
Department of Hematology, The Third Affiliated Hospital of Kunming Medical University, Kunming, China
Chengkui Geng
Department of Orthopedics, Yan’an Hospital of Kunming City, The Affiliated Hospital of Kunming Medical University, Kunming, China
Man Yang
Department of Endocrinology, The Affiliated Hospital of Yunnan University & The Second People’s Hospital of Yunnan Province, Kunming Yunnan, Kunming, Yunnan, China
Wenwen Mao
Department of Geriatics, The Second Hospital of Kunming, Kunming, China
Linlin Yang
Department of Gynecology, The Third Affiliated Hospital of Kunming Medical University, Kunming, China
Yin Ma
Department of Hematology, The Third Affiliated Hospital of Kunming Medical University, Kunming, China
Ming He
Department of Hematology, The Third Affiliated Hospital of Kunming Medical University, Kunming, China
Yeying Zhou
Department of Hematology, The Third Affiliated Hospital of Kunming Medical University, Kunming, China
Lianqing Liu
Department of Hematology, The Third Affiliated Hospital of Kunming Medical University, Kunming, China
Xuejiao Hu
Department of Hematology, The Third Affiliated Hospital of Kunming Medical University, Kunming, China
Jingxing Yu
Department of Hematology, The Second Affiliated Hospital of Kunming Medical University, Kunming, China
Xiufen Shen
Department of Laboratory, The Second Affiliated Hospital of Kunming Medical University, Kunming, China
Xuezhong Gu
Department of Hematology, The First People Hospital in Yunnan Province, Kunming, China
Liefen Yin
Department of Hematology, The Second Affiliated Hospital of Kunming Medical University, Kunming, China
Zhenglei Shen
Department of Hematology, The Third Affiliated Hospital of Kunming Medical University, Kunming, China
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.
