Thrombin induces IL-8/CXCL8 expression by DCLK1-dependent RhoA and YAP activation in human lung epithelial cells
Background: Doublecortin‑like kinase 1 (DCLK1) has been recognized as a marker of cancer stem cell in several malignancies. Thrombin is crucial in asthma severity as it can promote IL‑8/CXCL8 production in lung epithelial cells, which is a potent chemoattractant for neutrophils. However, the patho...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Springer Nature
2022
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| Online Access: | https://repository.ugm.ac.id/282842/1/209.pdf |
| _version_ | 1826050560025952256 |
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| author | Yuliani, Fara Silvia Chen, Jing-Yun Cheng, Wen-Hao Wen, Heng-Ching Chen, Bing-Chang Lin, Chien-Huang |
| author_facet | Yuliani, Fara Silvia Chen, Jing-Yun Cheng, Wen-Hao Wen, Heng-Ching Chen, Bing-Chang Lin, Chien-Huang |
| author_sort | Yuliani, Fara Silvia |
| collection | UGM |
| description | Background: Doublecortin‑like kinase 1 (DCLK1) has been recognized as a marker of cancer stem cell in several
malignancies. Thrombin is crucial in asthma severity as it can promote IL‑8/CXCL8 production in lung epithelial cells,
which is a potent chemoattractant for neutrophils. However, the pathologic role of DCLK1 in asthma and its involve‑
ment in thrombin‑stimulated IL‑8/CXCL8 expression remain unknown.
Methods: IL‑8/CXCL8, thrombin, and DCLK1 expression were observed in the lung tissues of severe asthma patients
and ovalbumin (OVA)‑induced asthmatic mice model. A549 and BEAS‑2B cells were either pretreated with inhibi‑
tors or small interfering RNAs (siRNAs) before being treated with thrombin. IL‑8/CXCL8 expression and the molecules
involved in signaling pathway were performed using ELISA, luciferase activity assay, Western blot, or ChIP assay.
Results: IL‑8/CXCL8, thrombin, and DCLK1 were overexpressed in the lung tissues of severe asthma patients and
ovalbumin (OVA)‑induced asthmatic mice model. Our in vitro study found that DCLK siRNA or LRKK2‑IN‑1 (DCLK1
inhibitor) attenuated IL‑8/CXCL8 release after thrombin induction in A549 and BEAS‑2B cells. Thrombin activated
DCLK1, RhoA, and YAP in a time‑dependent manner, in which DCLK1 siRNA inhibited RhoA and YAP activation. YAP
was dephosphorylated on the Ser127 site after thrombin stimulation, resulting in YAP translocation to the nucleus
from the cytosol. DCLK1, RhoA and YAP activation following thrombin stimulation were inhibited by U0126 (ERK
inhibitor). Moreover, DCLK1 and YAP siRNA inhibited κB‑luciferase activity. Thrombin stimulated the recruitment of YAP
and p65 to the NF‑κB site of the IL‑8/CXCL8 promoter and was inhibited by DCLK1 siRNA.
Conclusions: Thrombin activates the DCLK1/RhoA signaling pathway, which promotes YAP activation and transloca‑
tion to the nucleus from the cytosol, resulting in YAP/p65 formation, and binding to the NF‑κB site, which enhances
IL‑8/CXCL8 expression. DCLK1 might be essential in thrombin‑stimulated IL‑8/CXCL8 expression in asthmatic lungs
and indicates a potential therapeutic strategy for severe asthma treatment. |
| first_indexed | 2024-03-14T00:05:55Z |
| format | Article |
| id | oai:generic.eprints.org:282842 |
| institution | Universiti Gadjah Mada |
| language | English |
| last_indexed | 2024-03-14T00:05:55Z |
| publishDate | 2022 |
| publisher | Springer Nature |
| record_format | dspace |
| spelling | oai:generic.eprints.org:2828422023-11-17T01:07:59Z https://repository.ugm.ac.id/282842/ Thrombin induces IL-8/CXCL8 expression by DCLK1-dependent RhoA and YAP activation in human lung epithelial cells Yuliani, Fara Silvia Chen, Jing-Yun Cheng, Wen-Hao Wen, Heng-Ching Chen, Bing-Chang Lin, Chien-Huang Basic Pharmacology Background: Doublecortin‑like kinase 1 (DCLK1) has been recognized as a marker of cancer stem cell in several malignancies. Thrombin is crucial in asthma severity as it can promote IL‑8/CXCL8 production in lung epithelial cells, which is a potent chemoattractant for neutrophils. However, the pathologic role of DCLK1 in asthma and its involve‑ ment in thrombin‑stimulated IL‑8/CXCL8 expression remain unknown. Methods: IL‑8/CXCL8, thrombin, and DCLK1 expression were observed in the lung tissues of severe asthma patients and ovalbumin (OVA)‑induced asthmatic mice model. A549 and BEAS‑2B cells were either pretreated with inhibi‑ tors or small interfering RNAs (siRNAs) before being treated with thrombin. IL‑8/CXCL8 expression and the molecules involved in signaling pathway were performed using ELISA, luciferase activity assay, Western blot, or ChIP assay. Results: IL‑8/CXCL8, thrombin, and DCLK1 were overexpressed in the lung tissues of severe asthma patients and ovalbumin (OVA)‑induced asthmatic mice model. Our in vitro study found that DCLK siRNA or LRKK2‑IN‑1 (DCLK1 inhibitor) attenuated IL‑8/CXCL8 release after thrombin induction in A549 and BEAS‑2B cells. Thrombin activated DCLK1, RhoA, and YAP in a time‑dependent manner, in which DCLK1 siRNA inhibited RhoA and YAP activation. YAP was dephosphorylated on the Ser127 site after thrombin stimulation, resulting in YAP translocation to the nucleus from the cytosol. DCLK1, RhoA and YAP activation following thrombin stimulation were inhibited by U0126 (ERK inhibitor). Moreover, DCLK1 and YAP siRNA inhibited κB‑luciferase activity. Thrombin stimulated the recruitment of YAP and p65 to the NF‑κB site of the IL‑8/CXCL8 promoter and was inhibited by DCLK1 siRNA. Conclusions: Thrombin activates the DCLK1/RhoA signaling pathway, which promotes YAP activation and transloca‑ tion to the nucleus from the cytosol, resulting in YAP/p65 formation, and binding to the NF‑κB site, which enhances IL‑8/CXCL8 expression. DCLK1 might be essential in thrombin‑stimulated IL‑8/CXCL8 expression in asthmatic lungs and indicates a potential therapeutic strategy for severe asthma treatment. Springer Nature 2022 Article PeerReviewed application/pdf en https://repository.ugm.ac.id/282842/1/209.pdf Yuliani, Fara Silvia and Chen, Jing-Yun and Cheng, Wen-Hao and Wen, Heng-Ching and Chen, Bing-Chang and Lin, Chien-Huang (2022) Thrombin induces IL-8/CXCL8 expression by DCLK1-dependent RhoA and YAP activation in human lung epithelial cells. Journal of Biomedical Science, 29 (1). p. 95. ISSN 1423-0127 https://jbiomedsci.biomedcentral.com/articles/10.1186/s12929-022-00877-0 10.1186/s12929-022-00877-0 |
| spellingShingle | Basic Pharmacology Yuliani, Fara Silvia Chen, Jing-Yun Cheng, Wen-Hao Wen, Heng-Ching Chen, Bing-Chang Lin, Chien-Huang Thrombin induces IL-8/CXCL8 expression by DCLK1-dependent RhoA and YAP activation in human lung epithelial cells |
| title | Thrombin induces IL-8/CXCL8 expression by DCLK1-dependent RhoA and YAP activation in human lung epithelial cells |
| title_full | Thrombin induces IL-8/CXCL8 expression by DCLK1-dependent RhoA and YAP activation in human lung epithelial cells |
| title_fullStr | Thrombin induces IL-8/CXCL8 expression by DCLK1-dependent RhoA and YAP activation in human lung epithelial cells |
| title_full_unstemmed | Thrombin induces IL-8/CXCL8 expression by DCLK1-dependent RhoA and YAP activation in human lung epithelial cells |
| title_short | Thrombin induces IL-8/CXCL8 expression by DCLK1-dependent RhoA and YAP activation in human lung epithelial cells |
| title_sort | thrombin induces il 8 cxcl8 expression by dclk1 dependent rhoa and yap activation in human lung epithelial cells |
| topic | Basic Pharmacology |
| url | https://repository.ugm.ac.id/282842/1/209.pdf |
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