靶向肝癌细胞自噬提高大黄素的毒性杀伤作用

发布时间：2019-05-17 12:37

【摘要】：背景与目的:大黄素处理肝癌细胞后能够诱导内质网应激和凋亡。鉴于内质网应激与自噬之间的关联及后者作为细胞对抗应激环境的一种自我防御机制,该研究拟探讨通过抑制肝癌细胞自噬信号通路的策略提高大黄素对肿瘤细胞的毒性杀伤作用。方法:大黄素处理肝癌细胞后,应用CYTO-ID自噬检测试剂盒和蛋白[质]印迹法(Western blot)分别检测大黄素诱发细胞自噬情况;利用细胞自噬抑制剂(氯喹)预先抑制肝癌细胞自噬的产生,然后用大黄素处理肝癌细胞,最后通过ATPlite试验和细胞克隆形成实验检测肿瘤细胞存活;通过流式细胞术检测氯喹和大黄素联合处理诱导肝癌细胞发生凋亡的凋亡率,采用Western blot检测凋亡效应蛋白caspase-3活化断裂后产生活性片段的水平。结果:大黄素处理肝癌细胞后能够诱导肝癌细胞自噬;利用氯喹抑制肝癌细胞自噬能够显著能够提高大黄素对肝癌细胞克隆存活的抑制作用;氯喹和大黄素联合处理肝癌细胞能够显著提高细胞周期sub-G1期和活化caspase-3蛋白的表达水平。结论:靶向肝癌细胞自噬能够提高大黄素的毒性杀伤作用。
[Abstract]:Background & AIM: emodin can induce endoplasmic reticulum stress and apoptosis in HCC cells. In view of the relationship between endoplasmic reticulum stress and autophagy and the latter as a self-defense mechanism of cells against stress environment, The aim of this study was to investigate the toxic killing effect of emodin on tumor cells by inhibiting autophagy signaling pathway of HCC cells. Methods: after emodin treatment, CYTO-ID autophagy kit and protein printing (Western blot) were used to detect emodin induced autophagy. The autophagy inhibitor (chloroquine) was used to inhibit the autophagy of HCC cells in advance, and then the HCC cells were treated with emodin. Finally, the survival of HCC cells was detected by ATPlite test and cell clone formation assay. The apoptosis rate of HCC cells induced by combined treatment of chloroquine and emodin was detected by flow cytometry, and the level of active fragments produced by activation and fragmentation of apoptosis effect protein caspase-3 was detected by Western blot. Results: emodin could induce autophagy of HCC cells after treatment, and the inhibition of autophagy by chloroquine could significantly improve the inhibitory effect of emodin on the clone survival of HCC cells. Combined treatment of chloroquine and emodin could significantly increase the expression of sub- G 1 phase and activated caspase-3 protein in HCC cells. Conclusion: targeting autophagy of HCC cells can improve the toxic killing effect of emodin.
【作者单位】：甘肃省肿瘤医院腹外二科;
【基金】：甘肃省中医药管理局基金项目(GZK-2014-71)
【分类号】：R735.7

【相似文献】