Effect of P2X7R agonist BzATP on cell growth and apoptosis in non-small cell lung cancer A549 cells

Abstract

Abstract:ObjectiveTo investigate the expression of P2X7 receptor (P2X7R) and the effect of P2X7R agonist 2′3′O(4benzoylbenzoyl) ethane adenosine triphosphate three amine salt (BzATP) on cell growth and apoptosis in nonsmall cell lung cancer A549 cells, and to explore the related mechanism. MethodsThe expression of P2X7R in A549 cells was detected by immunofluorescence. Cells were treated with different concentrations (150, 300, 600 μmol/L) of BzATP. Cells untreated with BzATP were used as control group. 3(4,5dimethyl2thiazoly)2,5diphenyl2Htetrazolium bromide (MTT) assay and Hoest33342 staining were respectively used to detect cell viability and apoptosis. Enzymelinked immunosorbent assay (ELISA) was uesd to detect the concentration of tumor necrosis factorα (TNFα) of cell culture supernatants. The expressions of nuclear factorκB (NFκB) p65, inhibitor of α of NFκB (IκBα) and the phosphorylation of inhibitor of α of NFκB (phosphoIκBα) were detected by Western blotting. ResultsP2X7R was expressed on the cell membrane of A549 cells. Survival rate of A549 cell was significantly decreased with the concentrations of BzATP at 300 and 600 μmol/L ［(67.87±8.98)%, (44.73±6.92)%］, compared with the control group (98.60±1.44)%, the differences were statistically significant (t=4.481, P=0.027; t=3.920, P=0.038). BzATP promoted apoptosis, and increased the concentration of TNFα of supernatant at 300 and 600 μmol/L ［(57.35±6.41)pg/ml, (78.63±11.33)pg/ml］, compared with the control group (42.56±0.37)pg/ml, the differences were statistically significant (t=6.410, P=0.035; t=11.330, P=0.005). In addation, the expressions of NFκB p65 and IκBα were respectively downregulated and upregulated by BzATP, while the expression of phosphoIκBα was not significantly altered. ConclusionP2X7R is expressed on A549 cell membrane. BzATP can inhibit cell proliferation and induce the apoptosis of A549 cells, and the mechanism of action may be related to promoting the release of TNFα and inhibition of NFκB pathway.

Key words:Carcinoma, nonsmallcell lung,Apoptosis,P2X7 receptor,2′3′O(4benzoylbenzoyl) ethane adenosine triphosphate three amine salt

Zeng Kanghua, Ru Qin, Xiong Qi, Ai Yongxun. Effect of P2X7R agonist BzATP on cell growth and apoptosis in non-small cell lung cancer A549 cells[J]. Journal of International Oncology, 2016, 43(5): 321-325.

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