Xian Wei Su1, Gang Lu1, Chi Kwan Leung2, Qiang Liu1, Yi Li1, Kam Sze Tsang1, Shi Dou Zhao2, Danny Tat Ming Chan1,3, Hsiang Fu Kung4, Wai Sang Poon1,3. 1. Division of Neurosurgery, Department of Surgery, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, China. 2. Center for Reproductive Medicine, Shandong University, Jinan, China. 3. Otto Wong Brain Tumour Centre, Department of Surgery, Prince of Wales Hospital, The Chinese University of Hong Kong, Hong Kong, China. 4. School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong, China.
Abstract
OBJECTIVES: MicroRNAs (miRNAs) are considered as the cellular regulators which post-transcriptionally modulate gene expression in diverse biological processes including cell development and immunity. In this study, we investigated functions of miR-181d in dendritic cells (DCs) maturation, and the underlying mechanisms were also explored. MATERIALS AND METHODS: Here we did the miRNA screening in human DCs in response to lipopolysaccharides (LPS) by quantitative real-time PCR (qRT-PCR). The expressions of DCs maturation markers were measured after miRNA mimics transfections. The pharmacological inhibitors of signalling pathways were applied to examine miR-181d effect on DCs maturation by Western blot. Luciferase assay and mixed lymphocyte reaction (MLR) were also performed to reveal the target gene of miR-181d and test the viability of T cells treated with miR-181d transfected DCs. RESULTS: Overexpression of miR-181d per se is sufficient to promote DCs maturation, and up-regulate CD80 and CD83 expressions without LPS. Besides, we showed that miR-181d activated NF-κB pathway and also promoted the expression of pro-inflammatory cytokine IL12 and TNF-α. Inhibition of NF-κB pathway suppressed DCs maturation. Luciferase reporter assay and target gene knockdown assay indicated that miR-181d targets regulator cylindromatosis (CYLD), a primary negative regulator of NF-κB pathway. MLR assay showed that miR-181d-transfected DCs could promote T-cell proliferation than iDCs in vitro. CONCLUSION: Our study demonstrates that miR-181d is required for DCs maturation through the activation of NF-κB pathway by targeting CYLD.
OBJECTIVES: MicroRNAs (miRNAs) are considered as the cellular regulators which post-transcriptionally modulate gene expression in diverse biological processes including cell development and immunity. In this study, we investigated functions of miR-181d in dendritic cells (DCs) maturation, and the underlying mechanisms were also explored. MATERIALS AND METHODS: Here we did the miRNA screening in human DCs in response to lipopolysaccharides (LPS) by quantitative real-time PCR (qRT-PCR). The expressions of DCs maturation markers were measured after miRNA mimics transfections. The pharmacological inhibitors of signalling pathways were applied to examine miR-181d effect on DCs maturation by Western blot. Luciferase assay and mixed lymphocyte reaction (MLR) were also performed to reveal the target gene of miR-181d and test the viability of T cells treated with miR-181d transfected DCs. RESULTS: Overexpression of miR-181d per se is sufficient to promote DCs maturation, and up-regulate CD80 and CD83 expressions without LPS. Besides, we showed that miR-181d activated NF-κB pathway and also promoted the expression of pro-inflammatory cytokine IL12 and TNF-α. Inhibition of NF-κB pathway suppressed DCs maturation. Luciferase reporter assay and target gene knockdown assay indicated that miR-181d targets regulator cylindromatosis (CYLD), a primary negative regulator of NF-κB pathway. MLR assay showed that miR-181d-transfected DCs could promote T-cell proliferation than iDCs in vitro. CONCLUSION: Our study demonstrates that miR-181d is required for DCs maturation through the activation of NF-κB pathway by targeting CYLD.
Authors: Philipp Ströbel; Andreas Zettl; Zhou Ren; Petr Starostik; Hubertus Riedmiller; Stephan Störkel; Hans Konrad Müller-Hermelink; Alexander Marx Journal: Am J Surg Pathol Date: 2002-01 Impact factor: 6.394
Authors: Julien J Karrich; Loes C M Jachimowski; Marion Libouban; Anand Iyer; Kim Brandwijk; Esther W Taanman-Kueter; Maho Nagasawa; Esther C de Jong; Christel H Uittenbogaart; Bianca Blom Journal: Blood Date: 2013-09-06 Impact factor: 22.113
Authors: Xian Wei Su; Gang Lu; Chi Kwan Leung; Qiang Liu; Yi Li; Kam Sze Tsang; Shi Dou Zhao; Danny Tat Ming Chan; Hsiang Fu Kung; Wai Sang Poon Journal: Cell Prolif Date: 2017-07-21 Impact factor: 6.831