Role of Runt-related transcription factor 1 in regulating epithelial-mesenchymal transition in cigarette smoke extract stimulated rat airway epithelial cells_Chinese Journal of Respiratory and Critical Care Medicine

Authors：

HUANG Shuo ¹ , HU Ruicheng ² , TAN Shuangxiang ² , FU Daiyan ² ,  GAN Guixiang ²

1. Emergency Department, Hunan Provincial People's Hospital (The First Affiliated Hospital of Hunan Normal University), Changsha, Hunan 410016, P. R. China;
2. Department of Respiratory Medicine, Hunan Provincial People's Hospital (The First Affiliated Hospital of Hunan Normal University), Changsha, Hunan 410016, P. R. China;

Corresponding author：

GAN Guixiang, Email: ganguixiang2010@163.com

Keywords：

Epithelial-mesenchymal transition; Runt-related transcription factor 1; cigarette smoking extract; airway epithelial cells

DOI：

10.7507/1671-6205.202205037

Video：

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Objective To study the expression of human Runt-related transcription factor 1 (RUNX1) in rat airway epithelial cells stimulated by cigarette smoking extract (CSE), and explore the role of RUNX1 in regulating epithelial-mesenchymal transition (EMT). Methods Primary rat bronchial epithelial cells were cultured by enzyme digestion and stimulated with different concentrations of CSE. The viability of cells was detected by CCK-8 to explore the appropriate concentration of CSE. After the cells were treated with CSE, the Runx1 interference and overexpression vectors were constructed and transfected into the cells to silence or overexpress the Runx1 gene. Immunocytochemical method was used to detect RUNX1 expression and Western blot analysis was used to detect the expression of RUNX1, nuclear factor-κB (NF-κB), Snail, E-cadherin, and vimentin. Results The survival rate of bronchial epithelial cells could be reduced by CSE, and the degree of reduction was directly positively correlated to the concentration of CSE. After CSE stimulation, the expression level of E-cadherin in primary rat bronchial epithelial cells decreased significantly (P<0.05); the expression levels of RUNX1, NF-κB, Snail and vimentin significantly increased (P<0.05). After interfering with RUNX1 gene, the expression level of E-cadherin was up-regulated (P<0.05), and the expression levels of NF-κB, Snail and vimentin were down-regulated (P<0.05). After overexpression of RUNX1 gene, the expression level of E-cadherin decreased (P<0.05), and the expression levels of NF-κB, Snail and vimentin increased (P<0.05). Conclusions CSE promotes the expression of RUNX1 in rat airway epithelial cells. RUNX1 might regulate EMT process by involving in the regulation of NF-κB /Snail expression.

Citation： HUANG Shuo, HU Ruicheng, TAN Shuangxiang, FU Daiyan, GAN Guixiang. Role of Runt-related transcription factor 1 in regulating epithelial-mesenchymal transition in cigarette smoke extract stimulated rat airway epithelial cells. Chinese Journal of Respiratory and Critical Care Medicine, 2022, 21(9): 658-664. doi: 10.7507/1671-6205.202205037 Copy

1.	Jolly MK, Ward C, Eapen MS, et al. Epithelial-mesenchymal transition, a spectrum of states: role in lung development, homeostasis, and disease. Dev Dyn, 2018, 247(3): 346-358.
2.	Lee HW, Jose CC, Cuddapah S. Epithelial-mesenchymal transition: insights into nickel-induced lung diseases. Semin Cancer Biol, 2021, 76: 99-109.
3.	Sohal SS. Epithelial and endothelial cell plasticity in chronic obstructive pulmonary disease (COPD). Respir Investig, 2017, 55(2): 104-113.
4.	Eapen MS, Sharma P, Thompson IE, et al. Heparin-binding epidermal growth factor (HB-EGF) drives EMT in patients with COPD: implications for disease pathogenesis and novel therapies. Lab Invest, 2019, 99(2): 150-157.
5.	蔡琳, 吴壮, 徐军. 博莱霉素诱导α平滑肌肌动蛋白cre重组酶转基因小鼠肺纤维化上皮细胞-间质转分化的研究. 中国呼吸与危重监护杂志, 2009, 8(1): 52-56.
6.	邓玲玲, 欧阳博书, 魏颖, 等. 上皮间质转化在特发性肺纤维化及其信号通路中的研究进展. 复旦学报(医学版), 2022, 49(4): 614-619,27.
7.	Tang XJ, Sun L, Wang G, et al. RUNX1: a regulator of NF-κB signaling in pulmonary diseases. Curr Protein Pept Sci, 2018, 19(2): 172-178.
8.	Lin S, Zhang R, Xu L, et al. LncRNA Hoxaas3 promotes lung fibroblast activation and fibrosis by targeting miR-450b-5p to regulate Runx1. Cell Death Dis, 2020, 11(8): 706.
9.	林书典, 戴爱国, 席守民. 核因子-κb在大鼠慢性阻塞性肺疾病肺组织中的定位及表达. 中国应用生理学杂志, 2005, 21(3): 293-295.
10.	Vaquero J, Guedj N, Claperon A, et al. Epithelial-mesenchymal transition in cholangiocarcinoma: from clinical evidence to regulatory networks. J Hepatol, 2017, 66(2): 424-441.
11.	Tang XJ, Sun L, Jin XD, et al. Runt-related transcription factor 1 regulates LPS-induced acute lung injury via NF-κB signaling. Am J Respir Cell Mol Biol, 2017, 57(2): 174-183.
12.	Kiefel H, Bondong S, Pfeifer M, et al. EMT-associated up-regulation of L1CAM provides insights into L1CAM-mediated integrin signalling and NF-κB activation. Carcinogenesis, 2012, 33(10): 1919-1929.
13.	Zhu YW, Yan JK, Li JJ, et al. Knockdown of radixin suppresses gastric cancer metastasis in vitro by up-regulation of E-cadherin via NF-κB/snail pathway. Cell Physiol Biochem, 2016, 39(6): 2509-2521.
14.	Abs V, Bonicelli J, Kacza J, et al. Equine bronchial fibroblasts enhance proliferation and differentiation of primary equine bronchial epithelial cells co-cultured under air-liquid interface. PLoS One, 2019, 14(11): e0225025.
15.	McCarroll CS, He WH, Foote K, et al. Runx1 deficiency protects against adverse cardiac remodeling after myocardial infarction. Circulation, 2018, 137(1): 57-70.
16.	Hong DL, Fritz AJ, Gordon JA, et al. RUNX1-dependent mechanisms in biological control and dysregulation in cancer. J Cell Physiol, 2019, 234(6): 8597-609.
17.	Ramsey J, Butnor K, Peng Z, et al. Loss of RUNX1 is associated with aggressive lung adenocarcinomas. J Cell Physiol, 2018, 233(4): 3487-3497.
18.	裴银银, 黄丹怡, 张婷, 等. RUNX1在鼻息肉黏膜上皮细胞凋亡中的作用研究. 中华耳鼻咽喉头颈外科杂志, 2021, 56(12): 1328-1335.
19.	范文涛, 王攀红, 王倩. 马齿苋多糖对溃疡性结肠炎大鼠肠组织IL-6/STAT3及NF-κB的影响. 中国应用生理学杂志, 2018, 34(3): 263-267.
20.	黄云, 袁伟锋, 李理, 等. 谷胱甘肽硫转移酶M5通过p38MAPK/NF-κB途径下调肿瘤坏死因子α介导的人支气管上皮细胞炎症水平. 中国呼吸与危重监护杂志, 2016, 15(2): 171-175.
21.	Julien S, Puig I, Caretti E, et al. Activation of NF-κB by Akt upregulates snail expression and induces epithelium mesenchyme transition. Oncogene, 2007, 26(53): 7445-7456.

1. Jolly MK, Ward C, Eapen MS, et al. Epithelial-mesenchymal transition, a spectrum of states: role in lung development, homeostasis, and disease. Dev Dyn, 2018, 247(3): 346-358.
2. Lee HW, Jose CC, Cuddapah S. Epithelial-mesenchymal transition: insights into nickel-induced lung diseases. Semin Cancer Biol, 2021, 76: 99-109.
3. Sohal SS. Epithelial and endothelial cell plasticity in chronic obstructive pulmonary disease (COPD). Respir Investig, 2017, 55(2): 104-113.
4. Eapen MS, Sharma P, Thompson IE, et al. Heparin-binding epidermal growth factor (HB-EGF) drives EMT in patients with COPD: implications for disease pathogenesis and novel therapies. Lab Invest, 2019, 99(2): 150-157.
5. 蔡琳, 吴壮, 徐军. 博莱霉素诱导α平滑肌肌动蛋白cre重组酶转基因小鼠肺纤维化上皮细胞-间质转分化的研究. 中国呼吸与危重监护杂志, 2009, 8(1): 52-56.
6. 邓玲玲, 欧阳博书, 魏颖, 等. 上皮间质转化在特发性肺纤维化及其信号通路中的研究进展. 复旦学报(医学版), 2022, 49(4): 614-619,27.
7. Tang XJ, Sun L, Wang G, et al. RUNX1: a regulator of NF-κB signaling in pulmonary diseases. Curr Protein Pept Sci, 2018, 19(2): 172-178.
8. Lin S, Zhang R, Xu L, et al. LncRNA Hoxaas3 promotes lung fibroblast activation and fibrosis by targeting miR-450b-5p to regulate Runx1. Cell Death Dis, 2020, 11(8): 706.
9. 林书典, 戴爱国, 席守民. 核因子-κb在大鼠慢性阻塞性肺疾病肺组织中的定位及表达. 中国应用生理学杂志, 2005, 21(3): 293-295.
10. Vaquero J, Guedj N, Claperon A, et al. Epithelial-mesenchymal transition in cholangiocarcinoma: from clinical evidence to regulatory networks. J Hepatol, 2017, 66(2): 424-441.
11. Tang XJ, Sun L, Jin XD, et al. Runt-related transcription factor 1 regulates LPS-induced acute lung injury via NF-κB signaling. Am J Respir Cell Mol Biol, 2017, 57(2): 174-183.
12. Kiefel H, Bondong S, Pfeifer M, et al. EMT-associated up-regulation of L1CAM provides insights into L1CAM-mediated integrin signalling and NF-κB activation. Carcinogenesis, 2012, 33(10): 1919-1929.
13. Zhu YW, Yan JK, Li JJ, et al. Knockdown of radixin suppresses gastric cancer metastasis in vitro by up-regulation of E-cadherin via NF-κB/snail pathway. Cell Physiol Biochem, 2016, 39(6): 2509-2521.
14. Abs V, Bonicelli J, Kacza J, et al. Equine bronchial fibroblasts enhance proliferation and differentiation of primary equine bronchial epithelial cells co-cultured under air-liquid interface. PLoS One, 2019, 14(11): e0225025.
15. McCarroll CS, He WH, Foote K, et al. Runx1 deficiency protects against adverse cardiac remodeling after myocardial infarction. Circulation, 2018, 137(1): 57-70.
16. Hong DL, Fritz AJ, Gordon JA, et al. RUNX1-dependent mechanisms in biological control and dysregulation in cancer. J Cell Physiol, 2019, 234(6): 8597-609.
17. Ramsey J, Butnor K, Peng Z, et al. Loss of RUNX1 is associated with aggressive lung adenocarcinomas. J Cell Physiol, 2018, 233(4): 3487-3497.
18. 裴银银, 黄丹怡, 张婷, 等. RUNX1在鼻息肉黏膜上皮细胞凋亡中的作用研究. 中华耳鼻咽喉头颈外科杂志, 2021, 56(12): 1328-1335.
19. 范文涛, 王攀红, 王倩. 马齿苋多糖对溃疡性结肠炎大鼠肠组织IL-6/STAT3及NF-κB的影响. 中国应用生理学杂志, 2018, 34(3): 263-267.
20. 黄云, 袁伟锋, 李理, 等. 谷胱甘肽硫转移酶M5通过p38MAPK/NF-κB途径下调肿瘤坏死因子α介导的人支气管上皮细胞炎症水平. 中国呼吸与危重监护杂志, 2016, 15(2): 171-175.
21. Julien S, Puig I, Caretti E, et al. Activation of NF-κB by Akt upregulates snail expression and induces epithelium mesenchyme transition. Oncogene, 2007, 26(53): 7445-7456.

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Role of Runt-related transcription factor 1 in regulating epithelial-mesenchymal transition in cigarette smoke extract stimulated rat airway epithelial cells

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