Purinergic P2Y2 and P2X4 Receptors Are Involved in the Epithelial-Mesenchymal Transition and Metastatic Potential of Gastric Cancer Derived Cell Lines

Abstract

Gastric cancer (GC) is a major health concern worldwide, presenting a complex pathophysiology that has hindered many therapeutic efforts so far. In this context, purinergic signaling emerges as a promising pathway for intervention due to its known role in cancer cell proliferation and migration. In this work, we explored in more detail the role of purinergic signaling in GC with several experimental approaches. First, we measured extracellular ATP concentrations on GC-derived cell lines (AGS, MKN-45, and MKN-74), finding higher levels of extracellular ATP than those obtained for the non-tumoral gastric cell line GES-1. Next, we established the P2Y2 and P2X4 receptors (P2Y2R and P2X4R) expression profile on these cells and evaluated their role on cell proliferation and migration after applying overexpression and knockdown strategies. In general, a P2Y2R overexpression and P2X4R downregulation pattern were observed on GC cell lines, and when these patterns were modified, concomitant changes in cell viability were observed. These modifications on gene expression also modified transepithelial electrical resistance (TEER), showing that higher P2Y2R levels decreased TEER, and high P2X4R expression had the opposite effect, suggesting that P2Y2R and P2X4R activation could promote and suppress epithelial-mesenchymal transition (EMT), respectively. These effects were confirmed after treating AGS cells with UTP, a P2Y2R-agonist that modified the expression patterns towards mesenchymal markers. To further characterize the effects of P2Y2R activation on EMT, we used cDNA microarrays and observed that UTP induced important transcriptional changes on several cell processes like cell proliferation induction, apoptosis inhibition, cell differentiation induction, and cell adhesion reduction. These results suggest that purinergic signaling plays a complex role in GC pathophysiology, and changes in purinergic balance can trigger tumorigenesis in non-tumoral gastric cells.

ATP Promotes Oral Squamous Cell Carcinoma Cell Invasion and Migration by Activating the PI3K/AKT Pathway via the P2Y2-Src-EGFR Axis

“…In ovarian cancer, ATP induces EMT of ovarian cancer cells through the P2Y2 receptor-dependent activity of EGFR . In gastric cancer, purinergic P2Y2 and P2X4 receptors are involved in changes in the expression of EMT and related genes in gastric cancer cell lines . It is generally understood that ATP and other nucleotides, and their plasma membrane receptors play a central role in tumor cell proliferation and immune cell regulation.…”

“… 28 In gastric cancer, purinergic P2Y2 and P2X4 receptors are involved in changes in the expression of EMT and related genes in gastric cancer cell lines. 29 It is generally understood that ATP and other nucleotides, and their plasma membrane receptors play a central role in tumor cell proliferation and immune cell regulation. For this reason, an in-depth understanding of purinergic signals in the TME may provide new therapeutic prospects.…”

QSAR‐driven synthesis of antiproliferative chalcones against SH‐SY5Y cancer cells: Design, biological evaluation, and redesign

“…To assess the effects of chalcone derivatives on the expression of Bcl‐2 and Bax, plates were treated for 24 h with 15 µM chalcone 10b , using an untreated plate as basal expression condition, and 15 µM 5‐FU and CisP as positive controls (details in Supporting Information). [ 33 ]…”

Differential Distribution and Activity Profile of Acylpeptide Hydrolase in the Rat Seminiferous Epithelium

“…Acquired images were processed generating regions of interest (ROIs) using ZEN Imaging Software 3.4 (Carl Zeiss, Heidelberg, Germany), and figure composition was performed using Adobe Photoshop CS6 (Adobe Systems, San José, CA, USA). Signal colocalization was quantified using the Manders’ coefficients method as described previously [ 38 ]. To establish the area occupied by APEH (Alexa Fluor 488), we measured the pixel number of markers (APEH) that colocalized with acrosin signal and divided these values by the total pixels obtained for each marker.…”