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目的:探讨外源性三磷酸腺苷(ATP)对高表达P2X7受体的人宫颈癌Caski细胞(Caski-P2X7)生长及凋亡的影响,为临床应用提供理论依据。方法:选择本实验室建立的Caski-P2X7细胞和对照Caski-3.1细胞(转染空载体质粒),用不同浓度ATP分别处理两种细胞株,MTT法检测细胞生存率,荧光染色法检测线粒体膜电位,流式细胞术检测细胞凋亡率;用2.5mmol/L ATP处理两种细胞后,Western blot检测凋亡相关蛋白(细胞色素C,bcl-2,Bax)的变化。结果:不同浓度ATP作用后,两种细胞生存率及线粒体膜电位均降低,而细胞凋亡率升高,均呈浓度依赖性;同一浓度ATP作用后,Caski-P2X7细胞生存率及线粒体膜电位均显著低于Caski-3.1细胞(P<0.05),但细胞凋亡率显著高于Caski-3.1细胞(P<0.05)。2.5mmol/L ATP处理后,Caski-P2X7细胞胞质中细胞色素C及凋亡蛋白Bax表达量明显增加(P<0.05),而抗凋亡蛋白bcl-2表达量明显减少(P<0.05)。结论:一定浓度的ATP可通过P2X7受体途径诱导宫颈癌Caski细胞凋亡。
Objective: To investigate the effect of exogenous ATP on the growth and apoptosis of human cervical cancer Caski-P2X7 cells with high expression of P2X7 receptor, and to provide a theoretical basis for clinical application. Methods: Caski-P2X7 cells and control Caski-3.1 cells (transfected with empty vector plasmid) were selected in our laboratory. Two cell lines were treated with different concentrations of ATP. Cell viability was detected by MTT assay. Mitochondrial membrane The apoptosis rate was detected by flow cytometry. The apoptosis-related proteins (cytochrome C, bcl-2, Bax) were detected by Western blot after treated with 2.5 mmol / L ATP. Results: After treated with different concentrations of ATP, the survival rate of both cells and mitochondrial membrane potential decreased, while the apoptosis rate increased in a concentration-dependent manner. After treated with ATP at the same concentration, the survival rate of Caski-P2X7 cells and mitochondrial membrane potential Were significantly lower than Caski-3.1 cells (P <0.05), but the apoptosis rate was significantly higher than Caski-3.1 cells (P <0.05). After treatment with 2.5 mmol / L ATP, the expression of cytochrome C and apoptosis protein Bax in Caski-P2X7 cells increased significantly (P <0.05), while the expression of anti-apoptotic protein bcl-2 decreased significantly (P <0.05) . Conclusion: A certain concentration of ATP can induce cervical cancer Caski cell apoptosis through P2X7 receptor pathway.