雷公藤内酯酮通过调控Wnt/β-catenin信号通路诱导三阴性乳腺癌细胞凋亡与自噬  

Triptonide induces apoptosis and autophagy in triple-negative breast cancer cells by regulating Wnt/β-catenin signaling pathway

作　　者：吕仙梅 郭秋生 LYU Xianmei;GUO Qiusheng(Department of Radiotherapy,Jinhua People's Hospital,Jinhua,Zhejiang 321000,China;Department of Medical Oncology,Affiliated Jinhua Hospital,Zhejiang University School of Medicine,Jinhua,Zhejiang 321000,China)

机构地区：[1]浙江省金华市人民医院放疗科,金华321000 [2]浙江大学医学院附属金华医院肿瘤内科,金华321000

出　　处：《浙江中西医结合杂志》2024年第12期1106-1111,共6页Zhejiang Journal of Integrated Traditional Chinese and Western Medicine

基　　金：金华市中医药科技计划立项项目(No.2024LC11)。

摘　　要：目的研究雷公藤内酯酮(Tpn)对人三阴性乳腺癌(TNBC)细胞凋亡和自噬的诱导作用及其分子机制。方法采用CCK-8、克隆形成、Transwell、流式细胞术等实验分析Tpn对TNBC细胞株MDA-MB-231和MDA-MB-453的增殖、侵袭、凋亡和自噬的影响;利用蛋白免疫印迹(WB)实验检测Tpn对MDA-MB-231和MDA-MB-453细胞中Wnt 3a、β-连环蛋白(β-catenin)、切割型半胱氨酸蛋白酶-3(Cle-Caspase-3)、切割型聚腺苷二磷酸核糖聚合酶降解产物(Cle-PARP)、Beclin-1、p62、微管相关蛋白1轻链3(LC-3)等蛋白质的影响。结果不同浓度(10、20、40 nmol/L)的Tpn能够抑制MDA-MB-231和MDA-MB-453细胞株的克隆形成能力(48 h抑制率分别为39.67%、60.33%、79.14%和44.33%、52.34%、77.67%)和侵袭能力(48 h抑制率分别为26.33%、33.67%、45.67%和9.33%、15.34%、66.33%),并能诱导细胞凋亡(48 h凋亡率分别为10.06%、12.66%、18.76%和8.70%、15.09%、19.18%)。分子机制实验结果表明,Tpn处理MDA-MB-231和MDA-MB-453细胞后,Wnt 3a、β-catenin、p62等蛋白质的表达显著减少,而Cle-Caspase-3、Cle-PARP、Beclin-1等蛋白质的表达以及LC3Ⅱ/Ⅰ比值显著增加。结论Tpn通过调控Wnt/β-catenin信号通路抑制MDA-MB-231和MDA-MB-453细胞的克隆形成和侵袭能力,并诱导细胞凋亡和自噬。Objective To investigate the effects of triptonide(Tpn)on apoptosis and autophagy in human triple-negative breast cancer(TNBC)cells and the underlying molecular mechanisms.Methods The effects of Tpn on the proliferation,invasion,apoptosis,and autophagy of TNBC cell lines MDA-MB-231 and MDA-MB-453 were analyzed using CCK-8,colony formation,Transwell,and flow cytometry assays.Western blotting was utilized to de-tect the impact of Tpn on the expression of proteins including Wnt 3a,β-catenin,cleaved caspase-3(Cle-Caspa-se-3),cleaved PARP(Cle-PARP),Beclin-1,p62,and LC3 in MDA-MB-231 and MDA-MB-453 cells.Results Different concentrations(10,20,40 nmol/L)of Tpn were able to inhibit the colony formation ability of MDA-MB-231 and MDA-MB-453 cell lines,with 48-hour inhibition rates of 39.67%,60.33%,and 79.14%for MDA-MB-231,and 44.33%,52.34%,and 77.67%for MDA-MB-453,respectively.Additionally,Tpn inhibited their invasive abilities,with 48-hour inhibition rates of 26.33%,33.67%,and 45.67%for MDA-MB-231,and 9.33%,15.34%,and 66.33%for MDA-MB-453,respectively.Tpn also induced cell apoptosis,with 48-hour apoptosis rates of 10.06%,12.66%,and 18.76%for MDA-MB-231,and 8.70%,15.09%,and 19.18%for MDA-MB-453,respectively.Molecu-lar mechanism experiments showed that Tpn treatment significantly decreased the expression levels of proteins such as Wnt 3a,β-catenin,and p62,while significantly increased the expression levels of proteins such as Cle-Caspase-3, Cle-PARP, and Beclin-1, as well as the LC3 Ⅱ/Ⅰ ratio, in MDA-MB-231 and MDA-MB-453 cells. Conclusion Tpn inhibits the colony formation and invasion of MDA-MB-231 and MDA-MB-453 cells, as well as inducing apoptosis and autophagy. This mechanism is likely achieved through the regulation of the Wnt/β-catenin signaling pathway.

关 键 词：三阴性乳腺癌 雷公藤内酯酮 凋亡 自噬 WNT/Β-CATENIN信号通路 

分 类 号：R285[医药卫生—中药学]