摘要
Objective To assess the effects of Yi Qi Jie Du Decoction(YQJDD)on nasopharyngeal carcinoma stem cells(NPC-SCs)and investigate the underlying mechanism.Methods NPC-SCs were collected in serum-free culture system and identified by the sphere formation assay.The effect of YQJDD on the proliferation of NPC-SCs was detected using the Cell Counting Kit-8 assay.The change of mitochondrial membrane potential(ΔΨm)in NPC-SCs following treatment with YQJDD was investigated by JC-1 staining.The levels of apoptosisassociated proteins were examined using western blot analysis.Results YQJDD significantly inhibited the proliferation of NPCSCs in dose-and time-dependent manners.JC-1 staining revealed that YQJDD lowered theΔΨm of NPC-SCs in a timedependent manner.After treatment with YQJDD,the expression of cleaved caspase-3,-7 and-9,cleaved poly-ADP ribose polymerase,P21 and P53 were increased,while the expression of Survivin in NPC-SCs was decreased.However,the expression of cleaved caspase-8 remained nearly unchanged.Conclusions YQJDD inhibits the growth and induces apoptosis of NPC-SCs via an intrinsic apoptosis pathway.
目的研究益气解毒方(YQJDD)水提物对鼻咽癌干细胞(NPC-SCs)增殖和凋亡的影响及机制。方法采用无血清悬浮培养法分离培养NPC-SCs,成球实验鉴定NPC-SCs;CCK-8法检测YQJDD对NPC-SCs细胞增殖的影响;JC-1染色法分析YQJDD对NPC-SCs线粒体膜电位的影响;Western blot检测凋亡相关蛋白(cleaved caspase 3、7、8、9、cleaved PARP、P21、P53、Survivin)的表达。结果YQJDD可抑制NPC-SCs的增殖,且随着药物处理时间和浓度的增加,抑制效果增强(P<0.05);YQJDD可降低NPC-SCs细胞线粒体膜电位,诱导细胞凋亡;YQJDD可提高NPC-SCs细胞中cleaved caspase 3、7、9、cleaved PARP、P21、P53的表达,降低Survivin的表达,但对cleaved caspase 8的表达没有影响。结论YQJDD可通过线粒体凋亡途径诱导NPC-SCs细胞凋亡,抑制细胞增殖。
基金
funding support from the National Natural Science Foundation of China (No. 81573721, No. 81874408 and No. 81973914)
the Natural Science Foundation of Hunan (No. 2017JJ3246 and No. 2019JJ40216)
Excellent Youth Fund of Hunan Provincial Education Department (No. 19B439)
