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

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.

Arecoline promotes Akt‐c‐Myc‐driven aerobic glycolysis in esophageal epithelial cells

“…However, the specific type of ubiquitin chain and E3 ubiquitin ligase that mediated c‐Myc degradation needs further exploration. Based on previous research findings, arecoline is a compound carcinogen that induces genetic mutations, DNA damage, and epigenetic abnormalities in somatic cells 45–48 . The changes in glycolytic pathways induced by arecoline in esophageal epithelial cells in our study may also involve genetic mutations or epigenetic abnormalities in esophageal epithelial cells.…”

“…Moreover, both auto-oxidation of areca nut polyphenols in saliva and nitrosation of areca alkaloids contribute to the generation of ROS that drive the malignant transformation of OSF. 41 Jayasooriya et al 44 [45][46][47][48] The changes in glycolytic pathways induced by arecoline in esophageal epithelial cells in our study may also involve genetic mutations or epigenetic abnormalities in esophageal epithelial cells. However, the potential changes in esophageal epithelial cells induced by arecoline require further exploration of the underlying molecular mechanisms through sequencing analysis.…”

Targeting histone deacetylase 9 represses fibrogenic phenotypes in buccal mucosal fibroblasts with arecoline stimulation

“… 31 Furthermore, their transcriptome analysis of BMFs also indicated that the ECM-related and Wnt signaling were the distinct core signaling pathways that responded to the chronic treatment with arecoline. 32 However, other studies have shown that Wnt5a-mediated Wnt signaling activity is negatively associated with HDACs activity. 33 , 34 Notably, the available ChIP-Seq data from 8 cell lines revealed that the proximal promoter of HDAC9 contains transcription factor binding sites of not only MEF2A and MEF2C but also Egr-1 and NF-κB.…”

“… 30 , 37 Moreover, the arecoline-induced transcriptome profile of BMFs reported in a recent study supports our findings, indicating that low-dose arecoline-induced differentially expressed genes in BMFs were involved in ECM remodeling. 32 However, this is a relatively short duration compared to the OSF pathogenesis progression that may not reflect the potential long-term consequences of arecoline. Therefore, it is necessary to investigate the long-term treatment with arecoline on BMFs, such as conducting a stimulation period of at least a month in vitro, as well as arecoline-induced OSF animal models.…”

A Critical Interpretive Synthesis of the Role of Arecoline in Oral Carcinogenesis: Is the Local Cholinergic Axis a Missing Link in Disease Pathophysiology?

“…Arecoline, an active carcinogenic alkaloid found in areca nut, has been recognized as an important factor in the pathogenesis of premalignant and malignant oral disorders [1], specifically, oral submucous fibrosis (OSF) and oral squamous cell carcinoma (OSCC). However, the precise molecular mechanisms underlying arecoline-induced OSF and OSCC development remain unclear [2,3].…”