Effect of PADI4 on the Expression of Inflammatory Cytokines During NB4 Cells Differentiation.
10.19746/j.cnki.issn.1009-2137.2021.04.008
- Author:
Qing-Wei GUO
1
;
Fu LI
1
;
Li SONG
1
;
Ya-Ping WANG
1
;
Xiao-Mei YANG
2
Author Information
1. Department of Oncology & Hematology, Qilu Children's Hospital of Shandong University, Jinan 250022, Shandong Province, China.
2. Department of Oncology & Hematology, Qilu Children's Hospital of Shandong University, Jinan 250022, Shandong Province, China,E-mail: xuexian2010@163.com.
- Publication Type:Journal Article
- MeSH:
Cell Differentiation;
Cell Line, Tumor;
Cytokines/metabolism*;
Granulocytes;
Humans;
Leukemia, Promyelocytic, Acute;
Protein-Arginine Deiminase Type 4/metabolism*;
Tretinoin/pharmacology*
- From:
Journal of Experimental Hematology
2021;29(4):1065-1070
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the expression of peptidylarginine deiminase 4 (PADI4) during the process of differentiation into granulocyte of NB4 cells induced by all-trans-retinoic acid (ATRA) and whether PADI4 is involved in the inflammatory cytokines expression.
METHODS:Granulocyte differentiation model of NB4 cells induced by ATRA was established. The cell morphology changes were observed by Wright-Giemsa staining. The expression of cell differentiation marker CD11b was analyzed by flow cytometry. The mRNA and protein expression of PADI4 was detected by RT-PCR and Western blot, respectively. The expression of tumor necrosis factor (TNF) α and interleukin (IL) 1β was analyzed by ELISA, and also examined with the knockdown of PADI4 expression by siRNA.
RESULTS:After NB4 cells induced by ATRA, the cytoplasm increased and the ratio of nuclear to cytoplasmic was reduced. Nuclear dented, and rod-shaped nucleus, lobulated phenomenon increased (P<0.05). Flow cytometry analysis results showed that the cell surface molecule CD11b expression increased (P<0.01). RT-PCR and Western blot showed the expression of PADI4 increased at both transcriptional and translational levels during the process of the differentiation. ELISA showed TNF-α and IL-1β secretion increased in differentiated macrophages, while they could be inhibited by PADI4-specific siRNA.
CONCLUSION:During the differentiation into granulocyte of NB4 cells induced by ATRA, PADI4 expression increased. Furthermore, PADI4 appeared to play a critical role in inflammatory cytokines secretion.
