Pannexin1通道在顺铂诱导睾丸癌I-10细胞凋亡中的作用

Role of pannexin 1 channels in cisplatin-induced apoptosis in I-10 cells

目的观察pannexin1通道在顺铂诱导睾丸癌I-10细胞凋亡中的作用及其机制。方法 MTT法检测细胞存活率;Annexin V/PI双染法和Hoechst 33258染色法分别检测细胞早期和晚期凋亡;化学发光法检测细胞外ATP浓度;ELISA法检测细胞内IP3含量。结果 MTT法显示pannexin1通道抑制剂CBX与顺铂合用组的细胞存活率高于单用顺铂组(P<0.01);Annexin V/PI双染法显示CBX与顺铂合用组的细胞早期凋亡率低于单用顺铂组(P<0.001);Hoechst 33258染色法显示CBX与顺铂合用组的细胞晚期凋亡率低于单用顺铂组(P<0.01);化学发光法表明CBX与顺铂合用组的细胞外ATP浓度低于单用顺铂组(P<0.05);ELISA法表明CBX与顺铂合用组的细胞内IP3浓度低于单用顺铂组(P<0.05)。结论 Pannexin1通道参与顺铂诱导睾丸癌I-10细胞的凋亡,其机制可能与介导ATP/IP3信号通路有关。

Objective To investigate the role of pannexin 1 channels in cisplatin-induced apoptosis in I-10 cells and the mechanisms. Methods MTT assay was used to assess the cytotoxicity of cisplatin（DDP） in I-10 cells. Annexin V/PI double staining and Hoechst 33258 fluorescence staining were employed to detect early-and late-stage apoptosis of the cells,respectively. Extracellular ATP level and intracellular IP3 level in the cells were detected using commercial detection kits.Results I-10 cells exposed to both CBX（a pannexin 1 channel inhibitor） and DDP showed a higher cell viability compared with the cells exposed to DDP alone（P〈0.01）. CBX significantly decreased cisplatin-induced early-stage apoptosis（P〈0.001） and latestage apoptosis（P〈0.01）, and cause obvious reductions in extracellular ATP and intracellular IP3 levels during cisplatininduced apoptosis（P〈0.05）. Conclusion Pannexin 1 channels participate in cisplatin-induced apoptosis in I-10 cells possibly through the ATP/IP3 pathway.