葫芦素B通过p53及mTOR信号通路调控骨肉瘤细胞的增殖、迁移、侵袭和凋亡  

Cucurbitacin B regulates the proliferation,migration,invasion and apoptosis of osteosarcoma cells via p53 and mTOR signaling pathways

作　　者：黄海 韩莹 张哲龙 倪志明 于洪波 夏俊杰 HUANG Hai;HAN Ying;ZHANG Zhelong;NI Zhiming;YU Hongbo;XIA Junjie(Department of Orthopaedic Surgery,the 903rd Hospital of Joint Logistic Support Force of People's Liberation Army,Zhejiang Hangzhou 310000,China;Department of Orthopaedic Surgery,Sir Run Run Shaw Hospital,Zhejiang University School of Medicine,Zhejiang Hangzhou 310000,China)

机构地区：[1]中国人民解放军联勤保障部队第九〇三医院骨科,浙江杭州310000 [2]浙江大学医学院附属邵逸夫医院骨科,浙江杭州310000

出　　处：《现代肿瘤医学》2024年第17期3220-3227,共8页Journal of Modern Oncology

摘　　要：目的:探究葫芦素B(cucurbitacin B,CuB)对骨肉瘤细胞增殖、迁移、侵袭和凋亡的影响及其机制。方法:不同浓度的CuB干预骨肉瘤细胞系MG63和U2OS,CCK-8实验检测CuB对细胞活力的影响;克隆形成实验检测细胞增殖能力;划痕实验和Transwell实验检测细胞的迁移和侵袭能力;流式细胞术检测细胞周期和凋亡情况;Western blot法检测细胞周期相关蛋白(CyclinB1、CDK1)、凋亡相关蛋白(Cleaved PARP、BCL-2)、上皮间质转化(epithelial-mesenchymal transformation,EMT)相关蛋白(E-cadherin、N-cadherin、Vimentin、MMP)、p53转录因子及磷脂酰肌醇激酶(phosphatidylinositol-3-hydroxykinase,PI3K)/蛋白激酶B(protein kinase B,AKT)/雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)信号通路相关蛋白的表达。结果:CuB抑制骨肉瘤细胞系MG63和U2OS的增殖、迁移和侵袭能力,抑制细胞的上皮间质转化过程,将细胞周期阻滞于G_(2)/M期,并促进细胞凋亡。此外,CuB上调了骨肉瘤细胞中p53转录因子的表达及其磷酸化,抑制了PI3K的表达及mTOR、AKT的磷酸化。结论:CuB抑制骨肉瘤细胞的增殖、迁移和侵袭,并促进细胞凋亡。这一作用可能与p53及PI3K/AKT/mTOR信号通路的调控有关。Objective:To explore the effect and mechanism of cucurbitacin B(CuB) on cell proliferation,migration,invasion and apoptosis in osteosarcoma.Methods:MG63 and U2OS cell lines were treated with different concentrations of CuB,then CCK-8 assay was used to detect the effect of CuB on cell viability.Clonal formation assay was used to detect cell proliferation.Scratch assay and Transwell assay were used to detect cell migration and invasion.Flow cytometry was used to detect cell cycle and apoptosis.Western blot was used to detect the protein expression levels of cell cycle related proteins(CyclinB1,CDK1),apoptosis-related proteins(Cleaved PARP,BCL-2),epithelial-mesenchymal transformation(EMT)-related proteins(E-cadherin,N-cadherin,Vimentin,MMP),p53 transcription factor and PI3K/AKT/mTOR signaling pathway related proteins.Results:CuB suppressed cell proliferation,migration and invasion in MG63 and U2OS cell lines.Moreover,CuB inhibited epithelial mesenchymal transformation,induced G_(2)/M cell cycle arrest and promoted cell apoptosis.In addition,CuB up-regulated the expression and phosphorylation of p53,as well as inhibiting the expression of PI3K and phosphorylation of mTOR and AKT.Conclusion:CuB inhibits proliferation,migration and invasion of osteosarcoma cells and promotes apoptosis.This effect may be related to the regulation of p53 and PI3K/AKT/mTOR signaling pathway.

关 键 词：骨肉瘤 葫芦素B MTOR信号通路 P53 

分 类 号：R738.1[医药卫生—肿瘤]