摘要
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 effects of extracellular adenosine-5＇-triphosphate（ATP） on proliferation and apoptosis of cervical cancer Caski cells with P2X7 receptor over-experssion.Methods：The Caski-P2X7 cells and Caski-3.1 control cells（transfected mock plasmids） with different concentrations of ATP which were transfected and identified in my laboratory ago.MTT assay was used to assess the viability of cells.The change in cell membrane potential was measured by fluorescence staining.The apoptotic rates of these cells were detected by flow cytometry.After treatment with 2.5mmol/L ATP,the levels of apoptosis-related proteins（Cytochrome C,bcl-2,Bax）of two cells were tested by Western blot.Results：Two kinds of cell survival rate and mitochondrialmembrane potential were decreased,apoptosis rate increased,due to the treatment of different ATP concentration,and showed a dose-dependent.Caski-P2X7 cells survival rate and mitochondrialmembrane potential were significantly lower than that in Caski-3.1 cells under ATP treatment（P0.05）,but apoptosis rate was dramatically higher than that in Caski-3.1 cells（P0.05）.After treatment with 2.5mmol/L ATP,the expressions of cytochrome C and apoptosis protein Bax were increased significantly in Caski-P2X7 cell cytoplasm,and the expression of anti-apoptotic protein bcl-2 was significantly decreased.Conclusion：A certain concentration of ATP could induce apoptosis of Caski cells through P2X7 receptor pathway.
出处
《现代妇产科进展》
CSCD
2012年第11期857-860,共4页
Progress in Obstetrics and Gynecology
基金
湖北省自然科学基金资助项目(No:2009CDZ024)
