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Knockdown of Notch Suppresses Epithelial-mesenchymal Transition and Induces Angiogenesis in Oral Submucous Fibrosis by Regulating TGF-β1

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Abstract

Oral submucous fibrosis (OSF) is a chronic disorder with a high malignant transformation rate. Epithelial-mesenchymal transition (EMT) and angiogenesis are key events in OSF. The Notch signaling plays an essential role in the pathogenesis of various fibrotic diseases, including OSF. Our study aimed to explore the effects of Notch on the EMT and angiogenesis processes during the development of OSF. The expression of Notch in OSF tissues versus normal buccal mucosa samples was compared. Arecoline was used to induce myofibroblast transdifferentiation of buccal mucosal fibroblasts (BMFs). Short hairpin RNA technique was used to knockdown Notch in BMFs. Pirfenidone and SRI-011381 were used to inhibit and activate the TGF-β1 signaling pathway in BMFs, respectively. The expression of Notch was markedly upregulated in OSF tissues and fibrotic BMFs. Knockdown of Notch significantly decreased the viability and promoted apoptosis in BMFs subjected to arecoline stimulation. Downregulation of Notch also significantly suppressed the EMT process, as shown by the reduction of N-cadherin and vimentin with concomitant upregulation of E-cadherin. In addition, knockdown of Notch upregulated VEGF and enhanced the angiogenic activity of fBMFs. Moreover, inhibition of TGF-β1 suppressed viability and EMT, promoted apoptosis, and induced angiogenesis of fBMFs, while activation of TGF-β1 significantly diminished the effects of Notch knockdown on fBMFs. Knockdown of Notch suppressed EMT and induced angiogenesis in OSF by regulating TGF-β1, suggesting that the Notch-TGF-β1 pathway may serve as a therapeutic intervention target for OSF.

Highlights

  • Notch is upregulated in oral submucous fibrosis tissues and fibrotic buccal mucosal fibroblasts (fBMFs).

  • Knockdown of Notch suppresses the viability and epithelial-mesenchymal transition, promotes apoptosis, and induces angiogenesis in fBMFs.

  • Activation of the transforming growth factor beta 1 signaling diminished the effects of Notch knockdown on fBMFs.

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Data Availability

The raw data supporting the conclusions of this manuscript will be made available by the corresponding author, without undue reservation, to any qualified researcher.

Abbreviations

OSF:

Oral submucosal fibrosis

BMFs:

Buccal mucosal fibroblasts

EMT:

Epithelial-mesenchymal transition

shRNA:

Short hairpin RNA

fBMFs:

Fibrotic BMFs

VEGF:

Vascular endothelial growth factors

qRT-PCR:

Quantitative reverse transcription PCR

TGF-β1:

Transforming growth factor beta 1

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Acknowledgements

Not applicable.

Funding

This study was supported by the Hainan Provincial Natural Science Foundation of China (Grant No.822RC828) and Research Projects of Educational Science of Hainan Medical University (Grant No. HYZD202213).

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Author notes

  1. Jinrong Wang, Liyan Yang and Jie Mei share the first authorship.

Authors and Affiliations

  1. Department of Stomatology, The First Affiliated Hospital of Hainan Medical University, Haikou, 570102, China

    Jinrong Wang, Liyan Yang, Jie Mei, Zhixin Li, Yuqi Huang, Honglan Sun, Kaiyue Zheng, Huifang Kuang & Wen Luo

  2. School of Stomatology, Hainan Medical University, Haikou, 571199, China

    Jinrong Wang, Liyan Yang, Jie Mei, Zhixin Li, Yuqi Huang, Honglan Sun, Kaiyue Zheng, Huifang Kuang & Wen Luo

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  1. Jinrong Wang
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  4. Zhixin Li
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Contributions

Guarantor of integrity of the entire study: WL. Study concepts: WL. Study design: WL. Definition of intellectual content: ZL. Literature research: JW. Clinical studies: ZL. Experimental studies: JW. Data acquisition: JM. Data analysis: JM. Statistical analysis: HK. Manuscript preparation: YH. Manuscript editing: HS. Manuscript review: WL.

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Correspondence to Wen Luo.

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This study was approved by the Ethics Review Committee of the First Affiliated Hospital of Hainan Medical University.

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Wang, J., Yang, L., Mei, J. et al. Knockdown of Notch Suppresses Epithelial-mesenchymal Transition and Induces Angiogenesis in Oral Submucous Fibrosis by Regulating TGF-β1. Biochem Genet 62, 1055–1069 (2024). https://doi.org/10.1007/s10528-023-10452-3

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