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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。
Objective To investigate whether HBSS starvation buffer can induce the apoptosis of lung cancer cells and its possible mechanism. Methods Hank’s Balanced Solution (HBSS) starvation buffer was applied to Lewis cells for 12 h and Annexin V / PI flow cytometry was used to detect apoptosis. Lewis cells were exposed to ROS for 30 min after HBSS treatment, and the expression of HMGB1 protein was detected by Western blot 4, 8 and 12 h later. In addition, Caspase-3 expression was detected by Caspase-3 activity assay kit after Lewis cells were stimulated by HBSS for 12 h. Results HBSS starvation buffer significantly increased the percentage of apoptotic cells in Lewis cells compared with control cells after 12 h. The results of flow cytometry showed that the expression of ROS increased significantly in Lewis cells treated with HBSS for 30 min, and the expression of HMGB1 increased in HBSS-treated Lewis cells 3 h after immunoblotting. The secretion of HMGB1 was detected in the supernatant of cells 6 h after HBSS treatment. In addition, Caspase-3 expression was also significantly increased after 12 h stimulation by HBSS, with statistical significance. Conclusion HBSS starvation buffer can induce the apoptosis of lung cancer cells and the apoptosis may depend on the ROS-HMGB1-Caspase-3 pathway.