P2X7R激动剂BzATP对非小细胞肺癌A549细胞生长和凋亡的影响  

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

作　　者：曾康华[1] 茹琴[1] 熊琪[1] 艾永循[2] 

机构地区：[1]江汉大学武汉生物医学研究院,430056 [2]江汉大学武汉生物医学院,430056

出　　处：《国际肿瘤学杂志》2016年第5期321-325,共5页Journal of International Oncology

基　　金：国家自然科学基金（81302203）

摘　　要：目的研究配体门控离子通道P2x7受体（P2X7R）在非小细胞肺癌A549细胞中的表达，观察P2X7R激动剂2’-3’-O-（4-苯甲酰-苯甲酰）腺苷三磷酸三乙烷胺盐（BzATP）对A549细胞生长及凋亡的影响，并探究相关作用机制。方法采用免疫荧光法检测P2X7R在A549细胞中的表达。用不同浓度的BzATP（150、300、600μmol／L）处理，未用BzATP干预的细胞作为对照组。采用四甲基偶氮唑蓝（MTF）和Hoest33342染色法分别检测细胞存活率与凋亡情况，酶联免疫吸附试验检测上清液中肿瘤坏死因子-α（TNF-α）的浓度，Westernblotting检测核转录因子-κB（NF-κB）065,NF-κB抑制因子α（IκBα）及磷酸化NF-KB抑制因子α（phospho-IκBα）蛋白的表达。结果P2X7R在A549细胞膜上表达。在300、600μmol／LB zATP作用下A549细胞存活率分别为（67．87±8．98）％、（44．73±6．92）％，较对照组（98．60±1．44）％明显下降，差异均具有统计学意义（t=4．481，P=0．027；t=3．920，P=0．038）。BzATP可促进细胞凋亡，并且300、600μmol／LBzATP可上调细胞培养上清中TNF-α浓度，分别为（57．35±6．41）pg／ml、（78．63±11．33）pg／ml，与对照组（42．56±0．37）pg／ml比较差异具有统计学意义（t=6．410，P=0．035；t=11．330，P=0．005）。此外，BzATP可下调NF-κB p65的表达，上调IκBα的表达，对phospho-IκBα的表达无明显作用。结论P2X7R表达于A549细胞膜，BzATP能够抑制细胞增殖，促进细胞凋亡，其作用机制可能与促进细胞中TNF-α的释放，抑制NF-κB通路有关。Objective To investigate the expression of P2X7 receptor （P2X7R） and the effect of P2XTR 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 3.549 cells, and to explore the related mechanism. Methods The expression of P2XTR 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 （ MTY ） 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-st （TNF-α） of cell culture super- natants. The expressions of nuclear factor-KB （NF-κB） p65, inhibitor of st of NF-KB （IκBα） and the phos- phorylation of inhibitor of st of NF-κB （phospho-IκBα） were detected by Western blotting. Results P2XTR 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 differenees 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. Conclusion P2X7R is expressed on A549 cell membrane. BzATP can inhibit cell prolifera

关 键 词：癌 非小细胞肺 细胞凋亡 P2X7受体 2'-3-O-(4-苯甲酰.苯甲酰)腺苷三磷酸三乙烷 胺盐 

分 类 号：R734.2[医药卫生—肿瘤]