Omethoate induces pharyngeal cancer cell proliferation and G1/S cell cycle progression by activation of Akt/GSK-3β/cyclin D1 signaling pathway
Introduction
As an economical and efficient insecticide and acaricide, omethoate is one of the most frequently used organophosphorus pesticides (OPs) around the world, especially in China. If exposed to humans, the omethoate can accumulate for a long period of time after entering the body, which becomes harmful to human health (Rong et al., 2015; Sieke et al., 2018). This compoundIt has been caused public concern due to its toxic effects on non-target organisms after application (Pan et al., 2015) and is currently on the list of “Priority Monitoring Pesticides” published by the Ministry of Environmental Protection of the People’s Republic of China. Omethoate has toxic effects on non-target organisms after application (Pan et al., 2015). The public are easy to expose to omethoate, as residual omethoate has been detected in vegetables, fruits, grains and tea (Hao et al., 2011; Pan et al., 2015; Zhang et al., 2014). In the process of its use and production, it can also cause acute and chronic harm to the health of a person who come into contact with it, while the effects fromits low-dose exposure may go unnoticed (Duan et al., 2017a). Although, acute organophosphorous poisoning incidents have been gradually reduced in recent years, the toxic effects on human health caused by low-dose exposure to omethoate is attracting the public’s attention. Accumulating studies have concluded that long-term, low-dose exposure to OPs shares a close relationship with human tumorigenesis, adverse reproductive outcomes, and neurological and neurobehavioral function abnormalities (Duan et al., 2017a; Weichenthal et al., 2010). Presently, however, there is no direct experimental evidence of omethoate in cancer development thus far.
Head and neck squamous cell carcinoma (HNSCC) represents 6% of all cancer cases worldwide (Mourad et al., 2017; Vigneswaran and Williams, 2014). HNSCC is often found in the pharynx and larynx (Rothenberg and Ellisen, 2012) and is thought to account for 350,000 cancer deaths worldwide per year, with males affected significantly more than females at a ratio of 4:1 (Ferlay et al., 2018, 2010). Furthermore, for individuals involved in the production and pesticide spraying of omethoate, the absence of personal protective equipment could make it easier to inhale the compound, which may also increase the incidence of HNSCC. Moreover, due to its rapid growth and infiltration, the prognosis of HNSCC is not optimistic. Combination of operative and nonoperative treatment not only affects the patient’s survival, but also greatly influences the patient’s quality of life (Bai et al., 2018). As the molecular mechanism of HNSCC is still poorly understood, it becomes urgent to study its pathogenesis and development. Uncontrolled cellular proliferation is the hallmark of cancer, and tumor cells typically acquire damaged genes that directly regulate the cell cycle (Sherr, 1996).
Cyclins are essential regulators of the cell cycle machinery. Among them, cyclin D1 is one of the most frequently altered cell cycle regulators in cancers. Deregulated function of cyclin D1, often resulting from overexpression of its protein level, has been documented in numerous human cancers (Balandaram et al., 2016; He et al., 2013). Cyclin D1 is a multifunctional oncoprotein that regulates the G1 to S phase transition by binding to CDK4 or CDK6 and phosphorylating pRb (Pestell, 2013; van Caloen and Machiels, 2019). Akt also plays an important role in the regulation of cell growth, survival, differentiation, migration and metabolism by phosphorylating a number of proteins (Hlobilkova et al., 2007; Choi et al., 2018). It has been shown that Akt phosphorylates glycogen synthase kinase-3β (GSK-3β) at Ser9 residue and subsequently inhibits its kinase activity (Qin et al., 2015). Normally, GSK-3β negatively regulates cyclin D1 through enhancing proteasomal degradation of cyclin D1 by phosphorylating its Thr286 and inhibiting cyclin D1 gene transcription by increasing degradation of β-catenin (Ye et al., 2013). Thus, Akt phosphorylation and inhibition of GSK-3β will lead to the upregulation of cyclin D1.
In this study, we demonstrated that exposure of omethoate induced HNSCC cell proliferation and G1/S transition. Mechanistically, omethoate activated Akt, which led to the inhibition of GSK-3β and upregulation of cyclin D1. Our findings might provide direct evidence of omethoate promoting HNSCC growth.
Section snippets
Chemicals
Omethoate (Product Number: 36181) and Akt1/2 kinase inhibitor (1,3-Dihydro-1-(1-((4-(6-phenyl-1H-imidazo[4,5-g]quinoxalin-7-yl)phenyl)methyl)-4-piperidinyl)-2H-benzimidazol-2-one trifluoroacetate salt hydrate) (Product Number: A6730) were purchased from Sigma–Aldrich (St. Louis, MO).
Cell culture
The FaDu (human pharyngeal squamous carcinoma cell) cell line was purchased from Cell Bank of the Chinese Academy of Sciences in Shanghai and cultured in DMEM medium (10-013-CVR, Corning) supplemented with 10% fetal
Omethoate stimulates proliferation of FaDu cells
FaDu cells were treated with various concentrations (0, 0.1, 1, 10, 50, 100, 200 and 500 nM) of omethoate for 24 h. CCK-8 assay revealed that the number of FaDu cells increased in a dose-dependent manner, with the 100 nM dose demonstrating the highest rate of proliferation (Fig. 1A). We then used the 100 nM dosage for our time course studies, where FaDu cells were exposed to 100 nM of omethoate for different durations (0, 12, 24, 48 and 72 h). The results showed that FaDu cell growth was
Discussion
Omethoate is a broad category of organophosphorous pesticides (OPs) with increasing domestic use (Duan et al., 2017a). In recent years, acute poisoning incidents caused by OPs have been reduced, while the chronic toxic effect on human health caused by low-dose exposure to OPs has attracted the public’s attention (Celik et al., 2011). Omethoate exposure affects different organs including the lung, gastrointestinal tract, liver, brain and cardiomyocytes (Wang et al., 2016; Yan et al., 2012), and
Declaration of Competing Interest
The authors have no conflicts to disclose.
Acknowledgements
This work was supported in part by the National Natural Science Foundation of China (No. 81873249, No. 81602010) and the Natural Science Foundation of Shandong Province (ZR2019MH058, ZR2016HP16).
References (40)
- et al.
Head and neck cancer
Lancet
(2008) - et al.
Ligand activation of peroxisome proliferator-activated receptor-beta/delta suppresses liver tumorigenesis in hepatitis B transgenic mice
Toxicology
(2016) - et al.
Cancer incidence and mortality patterns in Europe: estimates for 40 countries and 25 major cancers in 2018
Eur. J. Cancer
(2018) - et al.
Wogonin induced G1 cell cycle arrest by regulating Wnt/beta-catenin signaling pathway and inactivating CDK8 in human colorectal cancer carcinoma cells
Toxicology
(2013) - et al.
Head and neck squamous cell carcinoma: update on epidemiology, diagnosis, and treatment
Mayo Clin. Proc.
(2016) - et al.
Epidemiological trends of head and neck cancer in the United States: a SEER population study
J. Oral Maxillofac. Surg.
(2017) New roles of cyclin D1
Am. J. Pathol.
(2013)- et al.
Involvement of Akt/GSK3beta/CREB signaling pathway on chronic omethoate induced depressive-like behavior and improvement effects of combined lithium chloride and astaxanthin treatment
Neurosci. Lett.
(2017) - et al.
Tumor necrosis factor alpha stimulates proliferation of dental pulp stem cells via Akt/glycogen synthase kinase-3beta/cyclin D1 signaling pathway
J. Endod.
(2015) - et al.
Human papillomavirus-related oropharyngeal cancer
Ann. Oncol.
(2017)
Hexachlorobenzene alters cell cycle by regulating p27-cyclin E-CDK2 and c-Src-p27 protein complexes
Toxicol. Lett.
Epidemiologic trends in head and neck cancer and aids in diagnosis
Oral Maxillofac. Surg. Clin. North Am.
Effects of omethoate on liver insulin signaling in mice
Biomed. Environ. Sci.: BES
2,3,7,8-tetrachlorodibenzo-p-dioxin stimulates proliferation of HAPI microglia by affecting the Akt/GSK-3beta/cyclin D1 signaling pathway
Toxicol. Lett.
Does omethoate have the potential to cause insulin resistance?
Environ. Toxicol. Pharmacol.
Normal range of human red blood cell acetylcholinesterase activity
Drug Chem. Toxicol.
Effects of MAPK and PI3K pathways on PD-L1 expression in melanoma
Clin. Cancer Res.
Golgi integral membrane protein 4 manipulates cellular proliferation, apoptosis, and cell cycle in human head and neck cancer
Biosci. Rep.
Neurotoxic and immunotoxic effects of fenthion and omethoate on frogs at acute exposure
Toxicol. Ind. Health
Adenosine induces intrinsic apoptosis via the PI3K/Akt/mTOR signaling pathway in human pharyngeal squamous carcinoma FaDu cells
Oncol. Lett.
Cited by (14)
Kaempferol ameliorates pulmonary vascular remodeling in chronic hypoxia-induced pulmonary hypertension rats via regulating Akt-GSK3β-cyclin axis
2023, Toxicology and Applied PharmacologyA systematic review of mechanistic studies on the relationship between pesticide exposure and cancer induction
2022, Toxicology and Applied PharmacologyCitation Excerpt :Phosphorylation of AKT and inhibition of GSK-3β will result in the cyclin D1 gene upregulation. Cycline D-1 upregulation by OP pesticides exposure (omethoate) can induce cells' G1 to S phase transition and increase pharyngeal cancer risk (Huo et al., 2019). Endosulfan exposure increased the expression level of ATM-p53 and caused cell cycle G1 arrest.
A ratiometric electrochemical biosensor via alkaline phosphatase mediated dissolution of nano-MnO<inf>2</inf> and Ru(III) redox recycling for the determination of dimethoate
2022, Journal of Pharmaceutical and Biomedical AnalysisCitation Excerpt :After use, it can enter natural waters or remain in rice, vegetables, and fruits, thus causing potential harm to human health. Therefore, it has been listed as one of the hazardous substances for priority monitoring by China's environmental protection department [19]. Inspired by the aforementioned developments, a split-type ratiometric electrochemical biosensor for the detection of dimethoate was proposed based on enzyme activity inhibition principles.
Copper exposure induces hepatic G0/G1 cell-cycle arrest through suppressing the Ras/PI3K/Akt signaling pathway in mice
2021, Ecotoxicology and Environmental SafetyCitation Excerpt :Akt, as an executer of PI3K, was shown to inactivate the transcription factor FOXO3A by phosphorylating Ser253 of FOXO3A, as well as GSK3-β by phosphorylating Ser9 of GSK3-β (Schubert et al., 2018). Generally, GSK-3β exerts its negative regulatory effect on cyclin D by strengthening proteasomal degradation of cyclin D through Thr 286 phosphorylation and inhibition of cyclin D gene transcription following an increase in β-catenin degradation (Huo et al., 2019). FOXO3A can target and activate multiple genes such as p21 and p27, and repress the production of cyclin D (Hornsveld et al., 2020).
Role of regulatory miRNAs of the Wnt/ β-catenin signaling pathway in tumorigenesis of breast cancer
2020, GeneCitation Excerpt :Therefore, β-catenin proteins accumulated in the cytosol can transferred to the nucleus and regulate multiple downstream genes like c-Myc and cyclin D1 (Gustafson, 2017; Anastas and Moon, 2013). Cyclins, which are considered as key promoters of cellular proliferation, have a prominent role in cell cycle progression through inducing cyclin dependent kinase enzymes (Lin, 2000; Huo, 2019). Nowadays, it is well-known that any defect in regulation of β-catenin target genes may causes un-controlled cell cycle progression and development of several types of human tumors like breast, lymphoma, melanoma, colorectal, and prostate cancers (Zhao, 2014; Gwak, 2017; Casimiro, 2014; Prasad, 2009).