PMID- 28986953
OWN - NLM
STAT- MEDLINE
DCOM- 20180806
LR  - 20180806
IS  - 1095-8355 (Electronic)
IS  - 1065-6995 (Linking)
VI  - 42
IP  - 2
DP  - 2018 Feb
TI  - CRISPR-Cas9 mediated gene knockout in human coronary artery endothelial cells 
      reveals a pro-inflammatory role of TLR2.
PG  - 187-193
LID - 10.1002/cbin.10885 [doi]
AB  - Endothelial inflammatory responses promote the development and progression of 
      atherosclerosis. It was reported that Toll-like receptors 2 (TLR2) is associated 
      with endothelial inflammation. However, the effect of TLR2 on inflammatory 
      responses in human coronary artery endothelial cells (HCAECs) remains largely 
      unknown. Here, we tested the hypothesis that TLR2 can enhance inflammatory 
      reactions in HCAECs after stimulated by TLR2 agonist. First, we used CRISPR-Cas9 
      technology to knockout TLR2 gene in HCAECs. Then, TLR2-KO and wild type HCAECs 
      were treated with TLR2 agonist peptidoglycan (PGN). The expression levels of 
      intercellular cell adhesion molecule-1 (ICAM-1), interleukin-6 (IL-6), and 
      interleukin-8 (IL-8) were analyzed by real-time PCR, Western blot, and ELISA. The 
      expression status of myeloid differentiation primary response gene 88 (MyD88), 
      phosphorylated IRAK-1 (pIRAK-1) and phosphorylated NF-kappaB (pNF-kappaB) were detected 
      by Western blot. Our results show that after treated with TLR2 agonist, the 
      expression levels of ICAM-1, IL-6, and IL-8 were downregulated in TLR2-KO cells 
      compared to those of wild type cells. Further, Western blots of MyD88, pIRAK-1, 
      and pNF-kappaB show that the expression levels of these pro-inflammatory molecules 
      were much lower in TLR2-KO cells compared to that of wild type cells by 
      stimulating with TLR2 agonist. We suggest that TLR2 may affect inflammatory 
      reaction in HCAECs by introducing pro-inflammatory molecules like MyD88, pIRAK-1, 
      and pNF-kappaB.
CI  - (c) 2017 International Federation for Cell Biology.
FAU - Wang, Yingge
AU  - Wang Y
AD  - The Department of Neurology, Affiliated Hospital of Yangzhou University, Yangzhou 
      University, Yangzhou, China.
AD  - Research Center for Vascular Biology, College of Medicine, Yangzhou University, 
      Yangzhou, China.
AD  - Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for 
      Prevention and Treatment of Senile Disease, Yangzhou University, Yangzhou, China.
FAU - Chen, Lu
AU  - Chen L
AD  - Research Center for Vascular Biology, College of Medicine, Yangzhou University, 
      Yangzhou, China.
FAU - Tian, Zheng
AU  - Tian Z
AD  - The Department of Cardiology Affiliated Zhongshan Hospital of Dalian University, 
      Dalian, China.
FAU - Shen, Xueyi
AU  - Shen X
AD  - Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for 
      Prevention and Treatment of Senile Disease, Yangzhou University, Yangzhou, China.
FAU - Wang, Xiaohong
AU  - Wang X
AD  - Research Center for Vascular Biology, College of Medicine, Yangzhou University, 
      Yangzhou, China.
FAU - Wu, Honghai
AU  - Wu H
AD  - Research Center for Vascular Biology, College of Medicine, Yangzhou University, 
      Yangzhou, China.
AD  - Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for 
      Prevention and Treatment of Senile Disease, Yangzhou University, Yangzhou, China.
FAU - Wang, Yayi
AU  - Wang Y
AD  - Research Center for Vascular Biology, College of Medicine, Yangzhou University, 
      Yangzhou, China.
FAU - Zou, Jiayu
AU  - Zou J
AD  - Research Center for Vascular Biology, College of Medicine, Yangzhou University, 
      Yangzhou, China.
FAU - Liang, Jingyan
AU  - Liang J
AD  - Research Center for Vascular Biology, College of Medicine, Yangzhou University, 
      Yangzhou, China.
AD  - Jiangsu Key Laboratory of Integrated Traditional Chinese and Western Medicine for 
      Prevention and Treatment of Senile Disease, Yangzhou University, Yangzhou, China.
AD  - Jiangsu Co-Innovation Center for Prevention and Control of Important Animal 
      Infectious Disease and Zoonoses, Yangzhou University, Yangzhou, China.
LA  - eng
PT  - Journal Article
DEP - 20171109
PL  - England
TA  - Cell Biol Int
JT  - Cell biology international
JID - 9307129
RN  - 0 (Cytokines)
RN  - 0 (ICAM1 protein, human)
RN  - 0 (Inflammation Mediators)
RN  - 0 (Peptidoglycan)
RN  - 0 (TLR2 protein, human)
RN  - 0 (Toll-Like Receptor 2)
RN  - 126547-89-5 (Intercellular Adhesion Molecule-1)
SB  - IM
MH  - CRISPR-Cas Systems
MH  - Cells, Cultured
MH  - Coronary Vessels/*cytology
MH  - Cytokines/biosynthesis
MH  - Endothelial Cells/drug effects/*immunology/metabolism
MH  - Endothelium, Vascular/cytology/*immunology/metabolism
MH  - Gene Knockout Techniques
MH  - Humans
MH  - Inflammation Mediators/*metabolism
MH  - Intercellular Adhesion Molecule-1/metabolism
MH  - Peptidoglycan/pharmacology
MH  - Toll-Like Receptor 2/agonists/genetics/*physiology
OTO - NOTNLM
OT  - CRISPR-Cas9
OT  - TLR2
OT  - atherogenesis
OT  - coronary artery endothelial cell
OT  - inflammatory
EDAT- 2017/10/08 06:00
MHDA- 2018/08/07 06:00
CRDT- 2017/10/08 06:00
PHST- 2017/05/29 00:00 [received]
PHST- 2017/09/30 00:00 [accepted]
PHST- 2017/10/08 06:00 [pubmed]
PHST- 2018/08/07 06:00 [medline]
PHST- 2017/10/08 06:00 [entrez]
AID - 10.1002/cbin.10885 [doi]
PST - ppublish
SO  - Cell Biol Int. 2018 Feb;42(2):187-193. doi: 10.1002/cbin.10885. Epub 2017 Nov 9.