论文部分内容阅读
目的探讨内源性一氧化氮在TRAIL诱导的细胞凋亡中的作用,以期了解许多肿瘤组织表达诱导性一氧化氮合成酶的分子和细胞学机制。方法用短暂转染、稳定转染方法在人胚胎肾细胞HEK293中进行重组质粒载体转染,并对可诱导表达一氧化氮合成酶NOS2载体筛选稳定可被诱导的细胞克隆。一氧化氮表达通过ponastrone A试剂进行诱导表达实现。Caspase活性通过相应试剂盒进行检测,并用流式细胞仪检测细胞凋亡情况。结果成功构建了稳定可诱导表达一氧化氮合成酶的细胞克隆;内源性一氧化氮可提高TRAIL诱导的细胞凋亡率,与凋亡相关的caspase-8激酶活性亦显增加。结论内源性一氧化氮可提高由TRAIL诱导的细胞凋亡率。*
Objective To investigate the role of endogenous nitric oxide in TRAIL-induced apoptosis in order to understand the molecular and cytological mechanisms of inducible nitric oxide synthase (NOS) expression in many tumor tissues. Methods Recombinant plasmids were transfected in human embryonic kidney cells HEK293 by transient transfection and stable transfection. The stable and inducible cell clones were obtained by inducible expression of nitric oxide synthase NOS2 vector. Nitric oxide expression is induced by ponastrone A reagent. Caspase activity was detected by the corresponding kit, and apoptosis was detected by flow cytometry. Results The cell clone stably expressing inducible nitric oxide synthase was constructed successfully. Endogenous nitric oxide could increase the apoptosis rate induced by TRAIL and the apoptosis-related caspase-8 kinase activity. Conclusion Endogenous nitric oxide can increase the rate of apoptosis induced by TRAIL. *