Serum and glucocorticoid-regulated kinase 1 regulates transforming growth factor β1-connective tissue growth factor pathway in chronic rhinosinusitis
Serum/glucocorticoid-regulated kinase 1 (SGK1) has been identified as a crucial regulator in fibrotic disorders. Herein, we explored SGK1 role in tissue remodeling of chronic rhinosinusitis (CRS). Lentivirus was employed to generate an SGK1-overexpressing human bronchial epithelial cell (16HBE) line...
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| Published in | Clinical immunology (Orlando, Fla.) Vol. 234; p. 108895 |
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| Main Authors | , , , , , , , , , , , , |
| Format | Journal Article |
| Language | English |
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United States
Elsevier Inc
01.01.2022
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| Abstract | Serum/glucocorticoid-regulated kinase 1 (SGK1) has been identified as a crucial regulator in fibrotic disorders. Herein, we explored SGK1 role in tissue remodeling of chronic rhinosinusitis (CRS).
Lentivirus was employed to generate an SGK1-overexpressing human bronchial epithelial cell (16HBE) line. To screen SGK1 downstream genes, RNA sequencing was performed on SGK1-overexpressing and control cell lines. To determine protein and gene expression levels, immunohistochemistry, western blotting, and quantitative real-time polymerase chain reaction were employed. Correlation analysis was performed using mRNA expression levels of SGK1, transforming growth factor β1 (TGF-β1), and connective tissue growth factor (CTGF) derived from CRS mucosal tissue and GEO database. Gene set enrichment analysis was conducted using gene sets from Molecular Signatures Database. The severity of symptoms in CRS patients was assessed using the 22-Item Sinonasal Outcome Test.
SGK1 overexpression significantly increased the expression of connective tissue growth factor (CTGF) in 16HBE cells (P < 0.01). Consistently, CTGF protein level was considerably greater in mucosal tissue of CRS without nasal polyps (CRSsNP) than in CRS with nasal polyps (CRSwNP) (P < 0.05) or in control subjects (P < 0.01). TGF-β1 protein level was higher in mucosal tissue of CRSsNP patients than in CRSwNP patients (P < 0.001) or in the control group (P < 0.01). mRNA levels of SGK1 and CTGF (P < 0.05, r = 0.668; P = 0.001, r = 0.630), TGF-β1 and CTGF (P < 0.05, r = 0.560; P < 0.05, r = 0.420), as well as SGK1 and TGF-β1(P < 0.05, r = 0.612; P < 0.05, r = 0.524) were significantly correlated in CRS mucosal tissue and GSE36830 dataset, respectively. TGF-β1-induced upregulated genes were significantly enriched in SGK1 overexpression group. In vitro assays, TGF-β1 promoted SGK1 and CTGF expression in a concentration- and time-dependent manner. Administrating an SGK1 inhibitor, GSK650394, significantly inhibited TGF-β1-induced CTGF expression in 16HBE and dispersed primary nasal polyp cells.
TGF-β1 stimulation significantly increases SGK1 and CTGF expression. By regulating TGF-β1-CTGF pathway, SGK1 may participate in tissue remodeling in the pathological mechanism of CRS. |
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| AbstractList | Serum/glucocorticoid-regulated kinase 1 (SGK1) has been identified as a crucial regulator in fibrotic disorders. Herein, we explored SGK1 role in tissue remodeling of chronic rhinosinusitis (CRS).
Lentivirus was employed to generate an SGK1-overexpressing human bronchial epithelial cell (16HBE) line. To screen SGK1 downstream genes, RNA sequencing was performed on SGK1-overexpressing and control cell lines. To determine protein and gene expression levels, immunohistochemistry, western blotting, and quantitative real-time polymerase chain reaction were employed. Correlation analysis was performed using mRNA expression levels of SGK1, transforming growth factor β1 (TGF-β1), and connective tissue growth factor (CTGF) derived from CRS mucosal tissue and GEO database. Gene set enrichment analysis was conducted using gene sets from Molecular Signatures Database. The severity of symptoms in CRS patients was assessed using the 22-Item Sinonasal Outcome Test.
SGK1 overexpression significantly increased the expression of connective tissue growth factor (CTGF) in 16HBE cells (P < 0.01). Consistently, CTGF protein level was considerably greater in mucosal tissue of CRS without nasal polyps (CRSsNP) than in CRS with nasal polyps (CRSwNP) (P < 0.05) or in control subjects (P < 0.01). TGF-β1 protein level was higher in mucosal tissue of CRSsNP patients than in CRSwNP patients (P < 0.001) or in the control group (P < 0.01). mRNA levels of SGK1 and CTGF (P < 0.05, r = 0.668; P = 0.001, r = 0.630), TGF-β1 and CTGF (P < 0.05, r = 0.560; P < 0.05, r = 0.420), as well as SGK1 and TGF-β1(P < 0.05, r = 0.612; P < 0.05, r = 0.524) were significantly correlated in CRS mucosal tissue and GSE36830 dataset, respectively. TGF-β1-induced upregulated genes were significantly enriched in SGK1 overexpression group. In vitro assays, TGF-β1 promoted SGK1 and CTGF expression in a concentration- and time-dependent manner. Administrating an SGK1 inhibitor, GSK650394, significantly inhibited TGF-β1-induced CTGF expression in 16HBE and dispersed primary nasal polyp cells.
TGF-β1 stimulation significantly increases SGK1 and CTGF expression. By regulating TGF-β1-CTGF pathway, SGK1 may participate in tissue remodeling in the pathological mechanism of CRS. Serum/glucocorticoid-regulated kinase 1 (SGK1) has been identified as a crucial regulator in fibrotic disorders. Herein, we explored SGK1 role in tissue remodeling of chronic rhinosinusitis (CRS).PURPOSESerum/glucocorticoid-regulated kinase 1 (SGK1) has been identified as a crucial regulator in fibrotic disorders. Herein, we explored SGK1 role in tissue remodeling of chronic rhinosinusitis (CRS).Lentivirus was employed to generate an SGK1-overexpressing human bronchial epithelial cell (16HBE) line. To screen SGK1 downstream genes, RNA sequencing was performed on SGK1-overexpressing and control cell lines. To determine protein and gene expression levels, immunohistochemistry, western blotting, and quantitative real-time polymerase chain reaction were employed. Correlation analysis was performed using mRNA expression levels of SGK1, transforming growth factor β1 (TGF-β1), and connective tissue growth factor (CTGF) derived from CRS mucosal tissue and GEO database. Gene set enrichment analysis was conducted using gene sets from Molecular Signatures Database. The severity of symptoms in CRS patients was assessed using the 22-Item Sinonasal Outcome Test.METHODSLentivirus was employed to generate an SGK1-overexpressing human bronchial epithelial cell (16HBE) line. To screen SGK1 downstream genes, RNA sequencing was performed on SGK1-overexpressing and control cell lines. To determine protein and gene expression levels, immunohistochemistry, western blotting, and quantitative real-time polymerase chain reaction were employed. Correlation analysis was performed using mRNA expression levels of SGK1, transforming growth factor β1 (TGF-β1), and connective tissue growth factor (CTGF) derived from CRS mucosal tissue and GEO database. Gene set enrichment analysis was conducted using gene sets from Molecular Signatures Database. The severity of symptoms in CRS patients was assessed using the 22-Item Sinonasal Outcome Test.SGK1 overexpression significantly increased the expression of connective tissue growth factor (CTGF) in 16HBE cells (P < 0.01). Consistently, CTGF protein level was considerably greater in mucosal tissue of CRS without nasal polyps (CRSsNP) than in CRS with nasal polyps (CRSwNP) (P < 0.05) or in control subjects (P < 0.01). TGF-β1 protein level was higher in mucosal tissue of CRSsNP patients than in CRSwNP patients (P < 0.001) or in the control group (P < 0.01). mRNA levels of SGK1 and CTGF (P < 0.05, r = 0.668; P = 0.001, r = 0.630), TGF-β1 and CTGF (P < 0.05, r = 0.560; P < 0.05, r = 0.420), as well as SGK1 and TGF-β1(P < 0.05, r = 0.612; P < 0.05, r = 0.524) were significantly correlated in CRS mucosal tissue and GSE36830 dataset, respectively. TGF-β1-induced upregulated genes were significantly enriched in SGK1 overexpression group. In vitro assays, TGF-β1 promoted SGK1 and CTGF expression in a concentration- and time-dependent manner. Administrating an SGK1 inhibitor, GSK650394, significantly inhibited TGF-β1-induced CTGF expression in 16HBE and dispersed primary nasal polyp cells.RESULTSSGK1 overexpression significantly increased the expression of connective tissue growth factor (CTGF) in 16HBE cells (P < 0.01). Consistently, CTGF protein level was considerably greater in mucosal tissue of CRS without nasal polyps (CRSsNP) than in CRS with nasal polyps (CRSwNP) (P < 0.05) or in control subjects (P < 0.01). TGF-β1 protein level was higher in mucosal tissue of CRSsNP patients than in CRSwNP patients (P < 0.001) or in the control group (P < 0.01). mRNA levels of SGK1 and CTGF (P < 0.05, r = 0.668; P = 0.001, r = 0.630), TGF-β1 and CTGF (P < 0.05, r = 0.560; P < 0.05, r = 0.420), as well as SGK1 and TGF-β1(P < 0.05, r = 0.612; P < 0.05, r = 0.524) were significantly correlated in CRS mucosal tissue and GSE36830 dataset, respectively. TGF-β1-induced upregulated genes were significantly enriched in SGK1 overexpression group. In vitro assays, TGF-β1 promoted SGK1 and CTGF expression in a concentration- and time-dependent manner. Administrating an SGK1 inhibitor, GSK650394, significantly inhibited TGF-β1-induced CTGF expression in 16HBE and dispersed primary nasal polyp cells.TGF-β1 stimulation significantly increases SGK1 and CTGF expression. By regulating TGF-β1-CTGF pathway, SGK1 may participate in tissue remodeling in the pathological mechanism of CRS.CONCLUSIONSTGF-β1 stimulation significantly increases SGK1 and CTGF expression. By regulating TGF-β1-CTGF pathway, SGK1 may participate in tissue remodeling in the pathological mechanism of CRS. |
| ArticleNumber | 108895 |
| Author | Han, Miaomiao Wang, Dehui Wang, Huan Li, Huabin Zhang, Peiyuan Sun, Xicai Song, Xiaole Lai, Yuting Liu, Quan Hu, Guohong Hu, Li Jiang, Wenxiu Zhang, Jia |
| Author_xml | – sequence: 1 givenname: Yuting surname: Lai fullname: Lai, Yuting organization: Department of Otolaryngology-Head and Neck Surgery, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China – sequence: 2 givenname: Peiyuan surname: Zhang fullname: Zhang, Peiyuan email: zhangpeiyuan@sibs.ac.cn organization: CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, No. 320, Yueyang Road, Xuhui District, Shanghai, China – sequence: 3 givenname: Huan surname: Wang fullname: Wang, Huan organization: Department of Otolaryngology-Head and Neck Surgery, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China – sequence: 4 givenname: Li surname: Hu fullname: Hu, Li organization: Department of Clinical Laboratory, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China – sequence: 5 givenname: Xiaole surname: Song fullname: Song, Xiaole organization: Department of Otolaryngology-Head and Neck Surgery, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China – sequence: 6 givenname: Jia surname: Zhang fullname: Zhang, Jia organization: Department of Otolaryngology-Head and Neck Surgery, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China – sequence: 7 givenname: Wenxiu surname: Jiang fullname: Jiang, Wenxiu organization: Department of Otolaryngology-Head and Neck Surgery, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China – sequence: 8 givenname: Miaomiao surname: Han fullname: Han, Miaomiao organization: Department of Clinical Laboratory, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China – sequence: 9 givenname: Quan surname: Liu fullname: Liu, Quan organization: Department of Otolaryngology-Head and Neck Surgery, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China – sequence: 10 givenname: Guohong surname: Hu fullname: Hu, Guohong email: ghhu@sibs.ac.cn organization: CAS Key Laboratory of Tissue Microenvironment and Tumor, Shanghai Institute of Nutrition and Health, University of Chinese Academy of Sciences, Chinese Academy of Sciences, No. 320, Yueyang Road, Xuhui District, Shanghai, China – sequence: 11 givenname: Xicai surname: Sun fullname: Sun, Xicai organization: Department of Otolaryngology-Head and Neck Surgery, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China – sequence: 12 givenname: Huabin surname: Li fullname: Li, Huabin email: allergyli@163.com organization: Department of Otolaryngology-Head and Neck Surgery, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China – sequence: 13 givenname: Dehui surname: Wang fullname: Wang, Dehui email: wangdehuient@sina.com organization: Department of Otolaryngology-Head and Neck Surgery, Eye and ENT Hospital, Fudan University, No. 83, Fenyang Road, Xuhui District, Shanghai, China |
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| Keywords | TGF-β1 Chronic rhinosinusitis SGK1 Tissue remodeling CTGF |
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| Snippet | Serum/glucocorticoid-regulated kinase 1 (SGK1) has been identified as a crucial regulator in fibrotic disorders. Herein, we explored SGK1 role in tissue... |
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| SubjectTerms | Adult Cells, Cultured Chronic Disease Chronic rhinosinusitis Connective Tissue Growth Factor - analysis Connective Tissue Growth Factor - genetics Connective Tissue Growth Factor - physiology CTGF Female Humans Immediate-Early Proteins - genetics Immediate-Early Proteins - physiology Male Middle Aged Protein Serine-Threonine Kinases - genetics Protein Serine-Threonine Kinases - physiology Rhinitis - etiology Rhinitis - metabolism Severity of Illness Index SGK1 Signal Transduction - physiology Sinusitis - etiology Sinusitis - metabolism TGF-β1 Tissue remodeling Transforming Growth Factor beta1 - analysis Transforming Growth Factor beta1 - genetics Transforming Growth Factor beta1 - physiology |
| Title | Serum and glucocorticoid-regulated kinase 1 regulates transforming growth factor β1-connective tissue growth factor pathway in chronic rhinosinusitis |
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