Extracellular ATP Induced S-Phase Cell Cycle Arrest via P2Y Receptor-Activated ERK Signaling in Poorly Differentiated Oral Squamous Cell Carcinoma SAS Cells

Abstract

Extracellular ATP in the tumor microenvironment exhibits either pro- or antitumor effect via interaction with P2Y receptors, but the intracellular signaling and functional roles of P2Y receptors in oral squamous cell carcinoma (OSCC) are unclear. We aimed to study the effect of ATP on OSCC cell lines and the potential mechanisms involved. Through GEPIA dataset analysis, high expression levels of mRNA encoding P2Y receptors, the ATP-induced G protein-coupled receptors, were associated with better overall patient survival in head and neck squamous cell carcinoma. qPCR analysis showed that the poorly differentiated OSCC SAS cell line, had higher P2RY1 expression level compared to the well-differentiated H103 and H376 cell lines. Western blotting and flow cytometry analyses revealed that ATP phosphorylated ERK and elevated intracellular calcium signaling in all tested cell lines. A significant S-phase cell cycle arrest was observed in SAS, and preincubation with the MEK inhibitor PD0325901 reversed the ATP-induced S-phase arrest. We further demonstrated that ATP induced a slight reduction in cell count and colony formation yet significant apoptosis in SAS. Overall, we postulate that the ATP-induced S-phase arrest effect in SAS cells may be regulated through P2Y receptor-mediated ERK signaling, thus suggesting a potential antitumor effect of ATP via interaction with its distinct profile of P2Y receptors.

Extracellular Adenosine Triphosphate Binding to P2Y1 Receptors Prevents Glutamate-Induced Excitotoxicity: Involvement of Erk1/2 Signaling Pathway to Suppress Autophagy

“…A previous study showed that activated ATP-dependent potassium channels promoted glioma cell proliferation by upregulated Erk1/2 signaling pathway ( Huang et al, 2009 ), which supported our findings that extracellular ATP pretreatment via Erk1/2 signaling pathway promoted SH-SY5Y cell proliferation ( Supplementary Figure 5 ). Interestingly, extracellular ATP induced the S-phase cell cycle arrest of oral squamous cell carcinoma SAS cells via its binding to P2Y1 receptors-mediated Erk1/2 signaling ( Lau et al, 2021 ). This research showed the antitumor effects of extracellular ATP were through interaction with P2Y1 receptors to promote Erk1/2 signaling, which was opposite to its effects in SH-SY5Y cells shown in the current study.…”

The circadian regulation of extracellular ATP

“…The activation of purinergic receptors participates in the stem cell differentiation and in the neuronal function [19,20]. In the pathogenesis of colorectal cancer and oral squamous cell carcinoma, eATP mediated cancer cell migration and invasion by the signalregulated kinase and inflammatory protein [21][22][23][24]. The occurrence of cancer is increasingly demonstrated to associate with the chronic disruption of circadian clock [25].…”