BzATP

This study investigated the role of P2X7 receptor (P2X7R) in the proliferation and metastasis of gastric cancer (GC).

The functional role and possible mechanism of P2X7R in the progression of GC were investigated through in vitro and in vivo experiments.

The results showed that ATP and its analogue BzATP increased calcium current in AGS and HGC-27 cells, while P2X7R antagonists A438079 and AZD9056 decreased the ATP-induced calcium influx. Activation of P2X7R increased the glycogen accumulation in GC cells, enhanced the stress ability of actin fibers and cell morphology changes, and promoted the proliferation, migration and invasion of GC cells. Conversely, the application of A438079, AZD9056 or siP2X7R inhibited the proliferation, migration and invasion of GC cells. Moreover, activation of P2X7R increased the expression levels of EMT/metastasis related genes MMP-2, MMP-9, N-cadherin, Zeb1, Vimentin and Snail, hut decreased the E-cadherin expression. While A438069, AZD9056, LY294002 or siP2X7R reversed the expression of the above genes. Activation of P2X7R activated P13/AKT/GSK-3beta signaling to promote the proliferation, migration and invasion of GC. Additionally, in vivo experiments showed that ATP activated P2X7R to induce the growth of tumors.

Our conclusion is that activation of P2X7R promotes the proliferation, metastasis and EMT of GC cells by activating P13/AKT/GSK-3beta signaling, and indicates that P2X7R may become a new potential target for GC treatment.

The P2X4 receptor is an ATP-gated ion channel that was proposed as a novel drug target for neuropathic pain, neurodegenerative diseases, epilepsy, and cancer. Animal models are indispensable for elucidating the role of receptors in health and disease and for drug development. Here, we present a systematic analysis and comparison of P2X4 receptors of nine different species that are relevant for basic and applied research, human, monkey, dog, guinea pig, pig, rabbit, rat, mouse, and zebrafish. 1321N1 astrocytoma cell lines stably expressing the respective receptor were generated by retroviral transfection. Calcium influx assays were performed to assess the effects of structurally diverse P2X4 receptor agonists and antagonists with the aim of exploring species differences. Most agonists showed significantly reduced potency in mice, rats, and zebrafish in comparison to the human P2X4 receptor. The physiological agonist ATP was more potent at monkey, rabbit, guinea pig, and pig P2X4 receptors (0.0285-0.0594 μM) than at the human ortholog (0.269 μM) and least potent at the zebrafish receptor (3.27 mM). 2-MeS-ATP exhibited similar potency (0.290-4.50 μM) in all investigated P2X4 receptor species except zebrafish and is therefore a suitable tool compound for studying P2X4 receptors across species. The allosteric P2X4 receptor antagonists BX430 and 5-BDBD displayed notable species differences, while the indole derivative PSB-OR-2020 blocked all investigated mammalian species with high potency (0.696-6.32 nM), exhibiting only minor species differences. These data will be valuable to support future in vitro and, in particular, in vivo studies.