内源性一氧化氮提高TRAIL诱导的细胞凋亡率

ENDOGENOUS NITRIC OXIDE PROMOTES TRAIL-MEDIATED APOPTOSIS

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 explore the function of endogenous nitric oxide in TRAIL-induced cell apoptosis,and in order to understand the molecular and cellular mechanisms of inducible nitric oxide expression in many tumors.Methods Transient transfection and stable transfection are used for the reconstructed plasmid transfection in human embryonic kidney cells HEK293,inducible expression of nitric oxide has been identified and the stable expression clones has been selected by antibiotic G418 after induction by ponasterone.Caspases activity has been detected by the appropriate kits and the flow cytometry was used to analyze the cell apoptosis.Results The stable and inducible expressed NOS2 clones has been successfully selected and established,and the results shown that endogenous nitric oxide can increase TRAIL-induced apoptosis,meanwhile the activities of the apoptosis associated caspase-8,have been enhanced with the nitric oxide expression.Conclusion TRAIL-induced apoptosis has been enhanced by the endogenous nitric oxide expression.