Low-dose arecoline regulates distinct core signaling pathways in oral submucous fibrosis and oral squamous cell carcinoma

Zhenming Li; You Fu; Yuhua Hu; Yun Zhu; Longwei Hu; Chaoji Shi; Yi Zhang; Jianjun Zhang; Shanghui Zhou

doi:10.1186/s12903-023-02887-2

Low-dose arecoline regulates distinct core signaling pathways in oral submucous fibrosis and oral squamous cell carcinoma

BMC Oral Health. 2023 Mar 25;23(1):171. doi: 10.1186/s12903-023-02887-2.

Authors

Zhenming Li^#^{1

2

3

4

5

6}, You Fu^#^{1

2

3

4

5

6}, Yuhua Hu^{2

3

4

5

6

7}, Yun Zhu^{1

2

3

4

5

6}, Longwei Hu^{1

2

3

4

5

6}, Chaoji Shi^{1

2

3

4

5

6}, Yi Zhang^{2

3

4

5

6

8}, Jianjun Zhang^{1

2

3

4

5

6}, Shanghui Zhou^{9

10

11

12

13

14}

Affiliations

¹ Department of Oral & Maxillofacial - Head & Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China.
² College of Stomatology, Shanghai Jiao Tong University, Shanghai, 200011, China.
³ National Center for Stomatology, Shanghai, 200011, China.
⁴ National Clinical Research Center for Oral Diseases, Shanghai, 200011, China.
⁵ Shanghai Key Laboratory of Stomatology, Shanghai, 200011, China.
⁶ Shanghai Research Institute of Stomatology, Shanghai, 200011, China.
⁷ Department of Oral Pathology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China.
⁸ Department of Endodontics and Operative Dentistry, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China.
⁹ Department of Oral & Maxillofacial - Head & Neck Oncology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, 200011, China. zhoushanghui@shsmu.edu.cn.
¹⁰ College of Stomatology, Shanghai Jiao Tong University, Shanghai, 200011, China. zhoushanghui@shsmu.edu.cn.
¹¹ National Center for Stomatology, Shanghai, 200011, China. zhoushanghui@shsmu.edu.cn.
¹² National Clinical Research Center for Oral Diseases, Shanghai, 200011, China. zhoushanghui@shsmu.edu.cn.
¹³ Shanghai Key Laboratory of Stomatology, Shanghai, 200011, China. zhoushanghui@shsmu.edu.cn.
¹⁴ Shanghai Research Institute of Stomatology, Shanghai, 200011, China. zhoushanghui@shsmu.edu.cn.

^# Contributed equally.

Abstract

Background: Betel nut chewing plays a role in the pathogenesis of oral submucous fibrosis (OSF) and oral squamous cell carcinoma (OSCC). As the major active ingredient of the betel nut, the effect of arecoline and its underlying mechanism to OSF and OSCC pathogenesis remain unclear.

Methods: Next-generation sequencing-based transcriptome and dRRBS analysis were performed on OSF and OSCC cells under low-dose arecoline exposure. Functional analyses were performed to compare the different roles of arecoline during OSF and OSCC pathogenesis, and key genes were identified.

Results: In this study, we identified that low-dose arecoline promoted cell proliferation of both NFs and OSCC cells via the acceleration of cell cycle progression, while high-dose arecoline was cytotoxic to both NFs and OSCC cells. We performed for the first time the transcriptome and methylome landscapes of NFs and OSCC cells under low-dose arecoline exposure. We found distinct transcriptome and methylome profiles mediated by low-dose arecoline in OSF and OSCC cells, as well as specific genes and signaling pathways associated with metabolic disorders induced by low-dose arecoline exposure. Additionally, low-dose arecoline displayed different functions at different stages, participating in the modulation of the extracellular matrix via Wnt signaling in NFs and epigenetic regulation in OSCC cells. After exposure to low-dose arecoline, the node roles of FMOD in NFs and histone gene clusters in OSCC cells were found. Meanwhile, some key methylated genes induced by arecoline were also identified, like PTPRM and FOXD3 in NFs, SALL3 and IRF8 in OSCC cells, indicating early molecular events mediated by arecoline during OSF and OSCC pathogenesis.

Conclusions: This study elucidated the contribution of low-dose arecoline to OSF and OSCC pathogenesis and identified key molecular events that could be targeted for further functional studies and their potential as biomarkers.

Keywords: Arecoline; Malignant progression; OPMD; OSCC; OSF.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Arecoline / toxicity
Carcinoma, Squamous Cell* / pathology
Epigenesis, Genetic
Head and Neck Neoplasms* / genetics
Humans
Mouth Mucosa / pathology
Mouth Neoplasms* / pathology
Oral Submucous Fibrosis* / genetics
Oral Submucous Fibrosis* / pathology
Signal Transduction
Squamous Cell Carcinoma of Head and Neck / pathology

Substances

Arecoline