HMGB1参与HBSS诱导的Lewis细胞凋亡及其相关机制研究  

HMGB1 involves in HBSS- induced Lewis apoptosis and the potential mechanism

作　　者：孔凡芝[1] 苏兆亮[2] 倪萍[2] 徐鑫鑫[2] 张盼[2] 佘鹏[1] 许化溪[2] 

机构地区：[1]江苏大学附属人民医院口腔科,镇江212002 [2]江苏大学免疫学研究所,镇江212013

出　　处：《免疫学杂志》2014年第12期1065-1068,共4页Immunological Journal

基　　金：江苏省六大人才高峰项目(2013-WSN-002);江苏省临床医学专项(BL2012059);江苏省妇幼课题(F201350)

摘　　要：目的探讨HBSS饥饿缓冲液能否诱导肺癌细胞(Lewis)凋亡及可能的机制。方法 Hank′s平衡溶液(HBSS)饥饿缓冲液作用于Lewis细胞12 h后采用Annexin V/PI流式双染检测细胞凋亡。Lewis细胞在HBSS处理30 min后流式检测ROS的产生,4、8、12 h后通过免疫印迹检测HMGB1蛋白的表达。另外,HBSS刺激Lewis细胞12 h后采用Caspase-3活性检测试剂盒检测Caspase-3的表达。结果 HBSS饥饿缓冲液作用于Lewis细胞12 h后与对照组相比凋亡百分比明显增多。流式结果表明Lewis细胞在HBSS处理30 min后ROS表达量明显上升,免疫印迹检测发现HBSS作用Lewis细胞3 h后HMGB1表达增加,6 h后在细胞培养上清中检测到HMGB1的分泌。另外,HBSS刺激12 h后Caspase-3表达也显著增加,且有统计学意义。结论 HBSS饥饿缓冲液能诱导肺癌细胞(Lewis)凋亡,且凋亡可能依赖于ROS-HMGB1-Caspase-3通路。This study conducted to investigate whether HBSS starvation could induce Lewis apoptosis and the potential mechanism underlying. The apoptotic ratios of Lewis were determined by the Annexin V-FITC/PI apoptosis kit after Lewis nutrition depletion for 12 hours. Furthermore, after 30 min under HBSS starvation, ROS generation was detected. Western blot was used to detect HMGB1 in Lewis after pre-treation with HBSS for 4, 8, 12 hours, moreover,Caspase-3 expression was detected 12 hours after. Data showed 12 hours after HBSS treatment, the apoptotic ratios of Lewis were increased. Furthermore, after 30 min under HBSS, ROS generation was markedly increased. After 3 h under HBSS, HMGB1 protein was increased; and 6 hours after HBSS starvation, HMGB1 was detected in supernatant.At the same time Caspase-3 expression was also distinctly increased. Taken together, the apoptosis of Lewis could be induced by HBSS starvation, which may be associated with ROS-HMGB1-Caspase-3 signal pathway.

关 键 词：HBSS Lewis细胞 ROS HMGB1 

分 类 号：R734.2[医药卫生—肿瘤]