鸦胆子素A通过EMT信号诱导非小细胞肺癌细胞凋亡  

Bruceine A induces apoptosis of non-small cell lung cancer cells by EMT signaling axis

作　　者：第伍丹琲 王虹[1] 陶红艳[1] 武凡祺[1] DI WU Dan-bei;WANG Hong;TAO Hong-yan;WU Fan-qi(Respiratory Dept,the Second Hospital of Lanzhou University,Lanzhou 730030,China)

机构地区：[1]兰州大学第二医院呼吸科,甘肃兰州730030

出　　处：《中国药理学通报》2024年第8期1474-1481,共8页Chinese Pharmacological Bulletin

基　　金：甘肃省自然科学基金资助项目(No.22JR5RA983);兰州大学第二医院“萃英科技创新”计划资助项目(No.CY2023-BJ-04)。

摘　　要：目的探究鸦胆子素A对非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞凋亡和侵袭的影响及相关分子机制。方法CCK-8法和克隆形成实验分别检测鸦胆子素A对NSCLC细胞增殖和克隆形成影响;3D基质胶球侵袭和流式细胞术分别检测其对细胞侵袭和凋亡影响;Western blot检测上皮间质转化(epithelial-mesenchymal transition,EMT)和凋亡关键蛋白表达;荷瘤裸鼠体内验证鸦胆子素A抗肿瘤作用;在线数据库、分子对接,DARTS实验分别预测和筛选鸦胆子素A作用靶点。结果鸦胆子素A明显抑制NSCLC细胞增殖和克隆形成能力、诱导细胞凋亡、抑制细胞侵袭,上调c-caspase3、c-PARP、Bax、E-cadherin蛋白表达,下调Bcl-2、N-cadherin、Vimentin、Snail蛋白表达(P<0.05);裸鼠荷瘤实验表明,注射鸦胆子素A后肿瘤体积和质量均明显减小(P<0.05)。靶点预测、分子对接结合DARTS综合分析发现,鸦胆子素A可增加HSP90α蛋白稳定性。结论鸦胆子素A通过激活EMT信号引起NSCLC细胞EMT水平降低、诱导细胞凋亡,其可能是通过与HSP90α结合来调控细胞功能,发挥抗癌作用。Aim To investigate the effect of bruceine A on apoptosis and invasion of non-small cell lung cancer(NSCLC)cells and to explore its possible molecular mechanism.Methods CCK-8 and cell colony assay were used to detect the effect of bruceine A on proliferation and clone formation ability of NSCLC cells.3D matrigel drop invasion assay and flow cytometry were used to detect cell invasion ability and apoptosis rate.Western blot was used to detect expressions of epithelial-mesenchymal transition(EMT)key proteins and apoptosis key proteins.The anti-tumor effect of bruceine A was detected on A549 xenograft mice in vivo.Online databases,molecular docking methods and drug affinity responsive target stability(DARTS)were used to predict and screen the potential binding targets of bruceine A.Results Bruceine A significantly inhibited the proliferation and colony formation of NSCLC cells.Bruceine A significantly induced the apoptosis in NSCLC cells and inhibited cell invasion ability,significantly up-regulated the protein expressions of c-caspase3,c-PARP,Bax and E-cadherin,and significantly down-regulated the protein expressions of Bcl-2,N-cadherin,Vimentin and Snail(P<0.05).In the tumor-bearing mouse models,bruceine A significantly decreased the tumor volume and tumor weight(P<0.05).Target prediction,molecular docking and Western blot analysis of DARTS sample revealed that bruceine A could increase the stabilization of HSP90α.Conclusions Bruceine A reduces the EMT levels and induces apoptosis in NSCLC cells through the EMT signaling axis.Bruceine A may exert its anti-cancer effect by binding to HSP90α.

关 键 词：鸦胆子素A 非小细胞肺癌 上皮间质转化 凋亡 靶点预测 分子对接 

分 类 号：R-332[医药卫生] R284.1R319R329.25R734.2