蒌慈散结方对人三阴性乳腺癌细胞MDA-MB-231及BT549的生长抑制作用及其机制研究

Study on the inhibitory effect and mechanism of Louci Sanjie Recipe on the growth of human triplenegative breast cancer cell MDA-MB-231 and BT549

目的观察蒌慈散结方对三阴性乳腺癌细胞的干预效应,并探讨其发挥作用的可能靶点和机制。方法将不同类型三阴性乳腺癌细胞分别分为蒌慈散结方组及空白对照组,给予蒌慈散结方或PBS处理,观察药物干预后细胞形态变化,检测细胞活力、细胞凋亡情况,分析干预后细胞转录组及相关蛋白的表达变化。结果蒌慈散结方组乳腺癌细胞密度降低,细胞聚集,体积缩小,细胞碎片增多,细胞的正常边界消失,细胞质萎缩;与空白对照组相比,蒌慈散结方组三阴性乳腺癌细胞增殖减少,细胞凋亡增加(P<0.05)。转录组分析显示,蒌慈散结方干预后,差异基因富集于细胞因子相互作用、JAK/Stat等信号通路;Western Blot检测发现,蒌慈散结方干预后Stat3蛋白表达明显降低(P<0.05),Bim、Cleaved caspase-3蛋白表达显著增加(P<0.05)。结论蒌慈散结方可能通过下调Stat3蛋白,促进促凋亡蛋白Bim和cleaved caspase-3表达增加而诱导三阴性乳腺癌细胞凋亡。

Objective To observe the intervention effect of Louci Sanjie Recipe(LCSJ)on triple-negative breast cancer cells(TNBC)and explore the possible targets and mechanisms of its role.Methods Different types of triple-negative breast cancer cells were divided into two groups:LCSJ group and blank control group,and were treated with LCSJ or PBS.The morphological changes of cells were observed after drug intervention,the cell viability and apoptosis were detected,and the expression changes of cell transcription group and related proteins were analyzed after intervention.Results The density of breast cancer cells decreased,the cells showed a state of aggregation and shrinkage,the cell fragments increased,the normal boundaries of cells disappeared and the cytoplasm shrank in the LCSJ group;compared with the control group,it could inhibit the proliferation of triple-negative breast cancer cells;compared with the blank control,LCSJ could promote cell apoptosis(P<0.05).Transcriptome analysis showed that after LCSJ intervention,differential genes were enriched in cytokine interaction,JAK/Stat and other signaling pathways;Western Blot showed that the expression of Stat3 was reduced in LCSJ treated cells(P<0.05),however,the expression of both Bim and Cleaved caspase-3 was elevated in LCSJ treated cells.Conclusion LCSJ may induce apoptosis of triple-negative breast cancer cells by down-regulating Stat3 protein and increasing the expression of pro-apoptosis proteins Bim and Cleved caspase-3.